miR-27a基因多态性与乳腺癌易感性的Meta分析

基于外泌体miR-27a-3p探究疏肝益肾方治疗乳腺癌内分泌耐药患者的临床研究张冬妮;卢雯平;卓至丽【期刊名称】《北京中医药》【年(卷),期】2024(43)1【摘要】目的探究血浆外泌体miR-27a-3p及其靶基因BAK1血浆蛋白表达水平对内分泌耐药乳腺癌患者预后及治疗效果的评价作用;探究疏肝益肾方对乳腺癌内分泌耐药患者血浆外泌体miR-27a-3p、靶基因BAK1血浆蛋白的影响。

方法采用非随机对照的临床研究方法,纳入内分泌治疗(ET)耐药和ET敏感的乳腺癌患者各10例,观察2组患者血浆外泌体miR-27a-3p及其靶基因BAK1血浆蛋白表达的差异;ET耐药患者使用氟维斯群+CDK4/6抑制剂+疏肝益肾方加减治疗2周,比较治疗前后血浆外泌体miR-27a-3p、靶基因BAK1血浆蛋白水平、中医证候积分总分、生活质量评分的变化。

结果治疗前,ET耐药组患者血浆外泌体miR-27a-3p的表达低于ET敏感组患者(P>0.05);血浆BAK1蛋白显著低于ET敏感组患者(P<0.01);中医证候积分总分和生活质量评分高于ET敏感组(P<0.01)。

治疗后,ET耐药组血浆外泌体miR-27a-3p表达及BAK1蛋白与治疗前比较,差异无统计学意义(P>0.05);ET耐药组中医证候积分总分、生活质量评分较治疗前降低(P<0.05,P<0.01)。

结论血浆外泌体miR-27a-3p及其靶基因BAK1血浆蛋白表达水平是评价乳腺癌内分泌治疗反应的潜在标志物;疏肝益肾方可以改善内分泌耐药患者的生活质量和中医证候评分,可能通过调控血浆外泌体miR-27a-3p及其靶基因BAK1的水平发挥作用。

【总页数】6页(P34-39)【作者】张冬妮;卢雯平;卓至丽【作者单位】中国中医科学院广安门医院【正文语种】中文【中图分类】R73【相关文献】1.益肾调肝方治疗乳腺癌患者类更年期综合征肝肾阴虚证的临床研究2.益肾化痰方治疗乳腺癌骨转移患者重度疼痛的临床研究3.舒肝益肾方联合内分泌治疗激素依赖型乳腺癌患者的临床效果4.调脾疏肝益肾方联合化疗治疗三阴性乳腺癌术后复发患者的临床观察5.疏肝益肾方治疗乳腺癌内分泌耐药的临床观察及对外泌体miR-221的影响因版权原因，仅展示原文概要，查看原文内容请购买。

Hsa-mir-27a genetic variant contributes to gastric cancer susceptibility through affecting miR-27a and target gene expressionQingmin Sun,1,6,7Haijuan Gu,2,7Ying Zeng,3,7Yi Xia,3You Wang,1Yali Jing,4Li Yang5and Bin Wang3,61Department of Pharmacy,The Affiliated Wuxi Hospital for Maternal and Child Health Care of Nanjing Medical University,Wuxi,Jiangsu;2Department of Pharmacy,Nantong Tumor Hospital,Nantong,Jiangsu Province;3Department of Pharmacology,Nanjing Medical University,Nanjing,Jiangsu Province;4Department of Endocrinology,Drum Tower Clinical Medical College of Nanjing Medical University,Nanjing,Jiangsu Province;5Department of General Surgery,First Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu Province,China(Received April1,2010⁄Revised June7,2010⁄Accepted June25,2010⁄Accepted manuscript online July6,2010⁄Articlefirst published online July27,2010)Aberrant microRNA(miRNA)expression is presently proposed to correlate with various human cancers and common single-nucleo-tide polymorphisms(SNP)at miRNA genes can influence the mat-uration of miRNAs or miRNA-mediated transcriptional regulation. However,whether miRNAs SNP alter gastric cancer susceptibility is still unclear.Here we investigated the possible role of a com-mon A/G polymorphism(rs895819)within hsa-mir-27a in the development or progression of gastric cancer,and assessed the effect of rs895819on the expression of miR-27a and its target gene Zincfinger and BTB domain containing10(ZBTB10).In the present case-control study,we found that subjects with the vari-ant genotypes(AG+GG)showed a significantly increased risk of gastric cancer relative to AA carriers(adjusted odds ratio=1.48, 95%confidence interval1.06–2.05;P=0.019).The elevated risk was especially evident in older subjects(age>58years),men, nonsmokers and rural subjects.A significant association of hsa-mir-27a variant genotypes with lymph node metastasis was also observed.Further functional analyses indicated that variant geno-types might be responsible for elevated miR-27a levels and reduced ZBTB10mRNA.Moreover,an inverse correlation was found between ZBTB10and miR-27a levels.In conclusion,we were thefirst to show that a common polymorphism(rs895819) in hsa-mir-27a,by modulating miR-27a and ZBTB10levels,acted as an important factor of the gastric cancer susceptibility.(Cancer Sci2010;101:2241–2247)G astric cancer is one of the commonest malignant tumors inthe world.(1)Of all the treatment modalities,only surgical resection may offer an opportunity for long-term survival.How-ever,in most cases,curative resection of the tumor is impossible because of the advanced stage at the time of diagnosis.(2)In this regard,early detection of gastric cancer currently is the most important measure to decrease disease-associated mortality. Therefore,it appears very important tofind novel diagnosis bio-markers to improve patient prognosis.MicroRNAs(miRNAs),a class of small endogenous noncod-ing RNA,negatively regulate post-transcriptional gene expres-sion by directly cleaving target mRNA or by inhibiting their translation.(3)Although the underlying biological functions are not completely clear,it has been shown to play important roles in a variety of cellular processes including apoptosis,differentia-tion and cell proliferation.(4–6)Recent studies have identified that aberrant miRNAs expression correlated with various human cancers such as colon tumors,breast cancer,lung cancer,pan-creatic cancer and gastric cancer.(7)There is increasing evidence that single nucleotide polymor-phisms(SNP)or mutations could make a significant contribution to disease susceptibility and outcome.The high degree of phylogenetic conservation in miRNA sequences determines that the functional variation in miRNAs may influence various bio-logical processes.Therefore,a mutation or a SNP in miRNA genes might influence the transcription of primary miRNAs(pri-miRNAs),maturation of miRNAs,or miRNA-mediated tran-scriptional regulation.(8)Recently,one study has systematically identified323SNP in227known human miRNAs,and12SNP are located within the miRNA precursors.(9)One A⁄G polymor-phism(rs895819)was found in hsa-mir-27a,and it was located at position40relative to thefirst nucleotide.MicroRNA-27a (MiR-27a)is found on chromosome19,and has been shown to function as oncogenes in gastric adenocarcinoma by targeting prohibitin.(10)The latest studies also revealed that oncogenic miR-27a is a target for anticancer drugs.(11,12)Moreover,Mer-tens-Talcott et al.(13)reported that transfection of antisense miR-27a led to increased Zincfinger and BTB domain contain-ing10(ZBTB10)levels in MDA-MB-231cells.Similarly,Scott et al.revealed that miR-27a suppressed the expression of ZBTB10mRNA,which could potentially inhibit gastrin gene expression by interfering with Sp1activation.(14,15)Transcrip-tion factor Sp1expression and activation could contribute to gastric cancer cell survival,growth,and angiogenesis.(16)Given the importance of ZBTB10in the expression of Sp1,we hypothe-sized that miR-27a might play a role in gastric cancer develop-ment and progression by modifying the levels of ZBTB10 mRNA.To expand our knowledge regarding the new polymorphism and biological function of miR-27a,the current study was designed to determine whether the polymorphism was associ-ated with a risk of gastric cancer.We also investigated miR-27a and its target gene ZBTB10expression in consideration of the genotype.As expected,our study is thefirst to show that a com-mon polymorphism in hsa-mir-27a,by modulating miR-27a and its target gene ZBTB10levels,acted as an important factor in gastric cancer susceptibility.Materials and MethodsSubjects.This hospital-based case-control study comprised 304gastric cancer cases and304cancer-free controls.Cases were inpatients newly diagnosed with histologically confirmed gastric cancer,consecutively recruited at the Affiliated Hospital of Nanjing Medical University.Those with secondary or recur-rent malignancies were excluded.The controls were selected from patients hospitalized because of a variety of nonmalignant6To whom correspondence should be addressed.E-mail:binwang@;qingminsun@7These authors contributed equally to this work.doi:10.1111/j.1349-7006.2010.01667.x Cancer Sci|October2010|vol.101|no.10|2241–2247diseases.Those controls with a previous history of cancer or severe clinical symptoms were excluded.All subjects were interviewed to obtain information on age,gender,smoking sta-tus,residence(urban or rural),past medical history and medical treatment by questionnaires.Individuals who formerly or cur-rently smoked‡10cigarettes per day on average were defined as smokers.In addition,the following parameters were obtained from the pathological reports of the gastric cancer patients stud-ied:tumor,node,metastasis(TNM)staging,differentiation grade(World Health Organization classification)and tumor location.For miR-27a and ZBTB10expression,65tumor tissue sam-ples with histological evidence of primary gastric carcinoma were obtained from Tissue Bank of Affiliated Hospital of Nanj-ing Medical University.All tissues were snap-frozen in liquid nitrogen after surgical resection and stored at)80°C until RNA extraction.Detailed clinical features for each tissue donor, including age,sex,smoking status,clinical staging,residence, tumor location and survival times were collected.All subjects were genetically unrelated ethnic Han Chinese.The study was approved by the Nanjing Medical University Affiliated Hospital Ethics Committee and informed consent was obtained from all participants.Genotyping.Genomic DNA was isolated from the buffy coat of blood or tissue samples according to standard phenol⁄chloro-form extraction procedures,as described previously.(17)The SNP(rs895819)was analyzed by polymerase chain reaction (PCR)-restriction fragment length polymorphism.The182-bp DNA fragment containing the polymorphic site was amplified using two primers(forward5¢-GAACTTAGCCACTGTGAA-CACCACTTGG-3¢,and reverse5¢-TTGCTTCCTGTCA-CAAATCACATTG-3¢).The forward primer was modified at position47(underlined),thus creating a recognition site for the restriction endonuclease Dra III(CACNNN⁄GTG)in the pres-ence of the T allele at position40.The PCR reaction was per-formed in a total volume of20l L containing2l L10·PCR buffer(MBI Fermentas,Vilnius,Lithuania),1.625mM MgCl2, 0.15mM dNTPs,0.25l M each primer,200ng of genomic DNA and1.1U of Taq DNA polymerase(MBI Fermentas).The reaction mixtures were denatured at95°C for5min,followed by32cycles of95°C for25s,60°C for30s and72°C for30s, with afinal elongation at72°C for6min.Six l L of the PCR product was digested with10U of Dra III(New England Bio-Labs,Waltham,MA,USA)at37°C overnight.The genotypes were assessed as follows:a single182bp fragment for the CC (GG)genotype;two fragments of155and27bp for the TT (AA)genotype;and three fragments of182,155and27bp for the TC(AG)genotype.The resultant fragments were electro-phoresed on a3%agarose gel containing ethidium bromide,and bands were then visualized by ultraviolet transillumination (Fig.1A).In addition,10%of the samples were randomly selected for repeat assays,and the results were100%concor-dant.Genotyping was performed without knowledge of the sub-jects’case and control status.The PCR products of the SNP with different genotypes were selected and verified by direct sequencing using ABI3730DNA analyzer(Applied Biosys-tems,Foster City,CA,USA)(Fig.1).Quantitative RT-PCR.Total RNA from the frozen tissue sam-ple was extracted using Trizol RNA isolation reagent(Invitro-gen,Carlsbad,CA,USA)according to the manufacturer’s protocol.TaqMan microRNA assays(Applied Biosystems Inc.) were used to quantify miR-27a expression.Small nuclear RNA, U6B(Applied Biosystems Inc.),was considered as the normali-zation control.In brief,cDNA was synthesized from total RNA using gene-specific stem-loop RT primers.Reverse transcriptase reactions contained60ng of total RNA,50nmol⁄L stem-loop RT primer,0.25mM each of dNTP,1·RT buffer,5.33U⁄l L MultiScribe reverse transcriptase and0.38U⁄l L RNase inhibi-tor.The RT reaction mixtures were incubated at16°C for 30min,42°C for30min and85°C for5min and then held at )20°C.The real-time PCR reaction was performed in a total volume of10l L containing0.67l L of RT product, 0.5l L·20TaqMan microRNA assay mix,5l L TaqMan·2 universal PCR Master Mix.The PCR parameters were as fol-lows:one cycle of95°C for10min,then40cycles of95°C for 15s and60°C for1min.ZBTB10and TATA binding protein(TBP)(control)mRNA levels were examined by SYBR Green quantitative PCR.(13) Reactions were performed according to the manufacturer’s instructions using Power SYBR Green PCR Master Mix(A)(B)(C)(D)Fig.1.PCR-RFLP analysis and automatedsequencing of single-nucleotide polymorphism(SNP)rs895819in hsa-mir-27a.(A)Representativegel electropherogram showing three genotypes ofSNP rs895819.The PCR products were digested withrestriction enzyme Dra III and analyzed on a3%agarose gel.M,DNA marker;lane1,TC(AG)heterozygous;lane2,TT(AA)homozygous;lanes3and4,CC(GG)homozygous.(B–D)DNA sequencingchromatograms of the PCR products confirmedrs895819polymorphism.Double peaks labeled withY and an arrow represented the heterozygousgenotype TC(AG).2242doi:10.1111/j.1349-7006.2010.01667.x(Applied Biosystems Inc.).Forward(F)and reverse(R)primers were used as follows:ZBTB10,GCTGGATAGTAGT-TATGTTGC and CTGAGTGGTTTGATGGACAGA;TBP, TGCACAGGAGCCAAGAGTGAA and CACATCACAGCTC-CCCACCA.All real-time reactions were conducted with the ABI Prism7300sequence detection system(Applied Biosys-tems Inc.).Relative expression was calculated using the cycle threshold(Ct)values supplied by the manufacturer.(18) Statistics.Quantitative variables departing from the normal distribution including age were summarized as the median and analyzed by Mann–Whitney rank sum test.Distribution of categorical variables and genotype frequencies between gastric cancer cases and controls were compared by the Pear-son v2test.Hardy–Weinberg equilibrium was assessed for controls by a goodness-of-fit v2test.The association between the SNP(rs895819)and risk of gastric cancer was estimated by odds ratios(OR)and95%confidence intervals(CI).Car-riers of the genotype AA comprised the reference group.The crude OR was assessed by the Woolf approximation method. The adjusted OR was computed by the unconditional logistic regression method,with adjustment for age,sex,smoking sta-tus,hypertension,diabetes and residence.Effects of the genetic variation on miR-27a and ZBTB10expression were examined using ANOVA.Correlation between the expression levels of ZBTB10mRNA and miR-27a was analyzed using the Pearson correlation coefficient.All the analyses were car-ried out with the SPSS13.0(SPSS Inc.,Chicago,IL,USA) and were based on two-tailed probability.A P value of<0.05 was considered statistically significant.ResultsBaseline characteristics.Demographic characteristics of the study participants are summarized in Table1.The sex and age distributions were fairly identical among the case and control subjects,indicating that the matching of controls to cases was adequate.There were no statistically significant differences between the cases and control subjects in terms of smoking sta-tus,residence,history of hypertension and diabetes.Most of the cases were adenocarcinoma(98.03%).Patients with cancer of the gastric cardia and non-cardia were85and219,respectively. Among those297gastric cancers with available clinicopatholog-ical data,52,36,142and67were T1,T2,T3and T4,respec-tively;41,139and117were reported with well,moderate and poor differentiation,respectively.Positive lymph nodes were identified in190cases.SNP rs895819and risk of gastric cancer.As presented in Table2,the frequency of G alleles in the cases was39.97%, which was higher than in the controls(32.73%)(v2=6.88; P=0.009).The difference in rs895819genotype distributions between the cases and controls was statistically significant (v2=6.55;P=0.038).The genotype distributions of the SNP among the controls(v2=3.75;P=0.053)and the cases (v2=1.69;P=0.194)were in Hardy–Weinberg equilibrium. When the genotype AA was used as the reference group,the adjusted OR of the GG genotype associated with the risk of gas-tric cancer was1.69(95%CI1.04–2.75),after adjustment for age,sex,smoking status,hypertension,diabetes and residence. Subjects carrying the AG genotype were at a41%elevated risk for the gastric cancer(95%CI0.99–2.00).Moreover,subjects with the variant genotypes(AG+GG)showed an overall increased risk of gastric cancer relative to AA carriers(adjusted OR=1.48,95%CI1.06–2.05).Stratified analysis.Table3shows the results of stratified analyses by the median age of controls(58years),sex,smok-ing status and residence with the SNP(rs895819).A statisti-cally significant association between increased gastric cancer risk and rs895819variant genotypes was observed in older (age>58years)subjects(adjusted OR=1.74,95%CI 1.09–2.79),but not in younger(age£58years)subjects.In men,possession of the variant genotypes was associated with a 53%increased risk of gastric cancer(adjusted OR=1.53,95% CI1.05–2.22),while the association was not statistically signif-icant in women.The elevated risk associated with the variant genotypes tended to be more evident in non-smokers(adjusted OR=1.45,95%CI1.01–2.10)than in smokers.In statistical analyses stratified by residence,only a significant association was observed in rural subjects(adjusted OR=1.80,95%CI 1.10–2.94).Additionally,the effect of the SNP(rs895819)was evaluated with stratification by clinicopathological characters of the gas-tric cancer patients(Table4).It is observed that the SNP was significantly associated with lymph node metastasis(adjusted OR=1.67,95%CI1.02–2.74).However,we found no signifi-cant association of the SNP and the location of the primary can-cer,depth of tumor infiltration or grade of differentiation, respectively.Table1.Demographic informationCharacteristics Cases n(%)(n=304)Controls n(%)(n=304)PGender(male),n(%)228(75.00)228(75.00) 1.000Age†(years)59(51–66.5)58(50–66)0.861Weight†(kg)62.5(55–70)65(58–72)0.012Hypertension,n(%)54(17.76)59(19.41)0.602Diabetes,n(%)17(5.59)28(9.21)0.088Smoking,n(%)73(24.01)58(19.08)0.139Residence,n(%)Rural146(48.03)139(45.72)0.569Urban158(51.97)165(54.28)†Median(25th–75th percentiles).Table2.Distributions of the rs895819genotype in cases and controls,and risk rs895819genotypes estimates for the variantHsa-mir-27a Genotype Cases n(%)Controls†n(%)Crude OR(95%CI)PAdjusted OR‡(95%CI)POverall304304AA115(37.83)145(47.70) 1.00 1.00AG135(44.41)119(39.14) 1.43(1.01–2.02)0.043 1.41(0.99–2.00)0.056 GG54(17.76)40(13.16) 1.70(1.06–2.74)0.028 1.69(1.04–2.75)0.033 AG+GG189(62.17)159(52.30) 1.50(1.08–2.07)0.014 1.48(1.06–2.05)0.019 rs895819A allele365(60.03)409(67.27)rs895819G allele243(39.97)199(32.73)†Distributions of the rs895819genotypes was in Hardy–Weinberg equilibrium,v2=3.749,P=0.053.‡Adjusted for age,sex,smoking status, hypertension,residence and diabetes.CI,confidence interval;OR,odds ratio.Sun et al.Cancer Sci|October2010|vol.101|no.10|2243Effect of SNP rs895819on miR-27a expression.To determine whether the identified polymorphism of hsa-mir-27a SNP rs895819could affect miR-27a expression,we screened65 tumor tissues obtained from gastric cancer patients.Among the 65patients,25were of the rs895819AA genotype,30of the AG genotype and10of the GG genotype.There was a signifi-cant influence of genotype on miRNA expression.As shown in Figure2A,expression of miR-27a in the AG+GG cases com-pared with the AA cases was significantly different,relative Ct ratios were73.74%±4.34%versus78.98%±7.31% (P<0.001)(a lower Ct ratio represents a higher expression). The results showed that a common A⁄G polymorphism (rs895819)within the hsa-mir-27a sequence increased the amount of miR-27a from the variant genotypes compared with the A allele.Impact of SNP rs895819on miR-27a target gene ZBTB10.In order to investigate the functional consequences of impaired processing of miR-27a on its target genes,we measured ZBTB10 mRNA levels in65gastric tumor tissues.Expression of ZBTB10 in the AG+GG genotypes had significantly lower levels than the AA genotype.Subjects with the AG+GG genotypes dis-played higher Ct ratios compared with the AA carriers (107.56%±7.05%versus101.68%±7.16%;P=0.003).These data suggested that A⁄G polymorphism in hsa-mir-27a affected the expression of ZBTB10mRNA(Fig.2B).Correlation of miR-27a with ZBTB10in gastric carcinoma patients.To further examine the correlation between miR-27a and ZBTB10mRNA,we performed a Pearson correlation analy-sis.The result is shown in Fig.2C,and as expected,we found that ZBTB10mRNA expression inversely correlated with expression of miR-27a(r=)0.379;P=0.002).It suggests that ZBTB10was a direct and functional target of miR-27a.DiscussionIn the present study,we found that the common A⁄G polymor-phism within hsa-mir-27a(rs895819)was associated with a sig-nificantly increased risk of gastric cancer.Preliminary functional assays revealed the miR-polymorphism conferred a higher level of miR-27a,which accompanied significantly reduced ZBTB10mRNA.Therefore,we might have identified a genetic mechanism and a molecular phenotype underlying the involvement of miR-27a in the susceptibility for gastric cancer. Nowadays,a genome-wide association study is encouraging and widely conducted to help develop more accurate diagnostic and therapeutic strategies of various kinds of human tumors.(19) Our previous epidemiological observations also provided the evidence that the risk of gastric cancer was associated with genetic polymorphisms.(20–22)Recently,a class of novel func-tional polymorphisms in miRNAs and its binding sites are theTable3.Stratified analyses for variant rs895819genotypes in cases and controlsVariable(AG+GG)⁄AACrude OR(95%CI)P Adjusted OR†(95%CI)P Cases ControlsAge(median)£5884⁄6381⁄75 1.23(0.78–1.94)0.362 1.19(0.75–1.89)0.469 >58105⁄5278⁄70 1.81(1.14–2.88)0.012 1.74(1.09–2.79)0.021 GenderFemale50⁄2645⁄31 1.32(0.69–2.56)0.402 1.33(0.69–2.59)0.396 Male139⁄89114⁄114 1.56(1.08–2.27)0.019 1.53(1.05–2.22)0.027 Smoking statusSmokers49⁄2432⁄26 1.66(0.81–3.38)0.162 1.55(0.74–3.24)0.247 Non-smokers140⁄91127⁄119 1.44(1.00–2.07)0.048 1.45(1.01–2.10)0.045 ResidenceUrban91⁄6789⁄76 1.16(0.75–1.80)0.509 1.19(0.76–1.86)0.440 Rural98⁄4870⁄69 2.01(1.25–3.25)0.004 1.80(1.10–2.94)0.020†Adjusted for age,sex,smoking status,hypertension,residence and diabetes.CI,confidence interval;OR,odds ratio.Table4.Associations between variant rs895819genotypes and clinicopathological characteristics of gastric cancer†Variable(AG+GG)⁄AA Crude OR(95%CI)P Adjusted OR‡(95%CI)P Tumor differentiationWell251611Moderate8653 1.04(0.51–2.12)0.918 1.29(0.60–2.78)0.513 Poor7344 1.06(0.51–2.20)0.872 1.37(0.61–3.07)0.449 Depth of tumor infiltrationT1331911T219170.64(0.27–1.53)0.3160.71(0.27–1.86)0.486 T39250 1.06(0.55–2.05)0.864 1.02(0.51–2.01)0.965 T440270.85(0.40–1.80)0.6760.89(0.41–1.96)0.778 Lymph node metastasisNegative584911Positive12664 1.66(1.02–2.70)0.039 1.67(1.02–2.74)0.042 LocalizationCardia503511Non-cardia13980 1.22(0.73–2.03)0.453 1.33(0.78–2.29)0.297†Data for seven plaintively treated cases was not obtained for the inoperable tumors.‡Adjusted for age,sex,smoking status,hypertension, residence and diabetes.CI,confidence interval;OR,odds ratio.2244doi:10.1111/j.1349-7006.2010.01667.xmost interesting and powerful.MiRNAs,a class of novel post-transcriptional regulators,represent only a small part of the gen-ome,but they regulate almost one-third of human genes.(23) Several studies have shown that miRNA polymorphisms or aberrant expression is associated with various human can-cers.(24,25)In gastric carcinoma,miR-15b and miR-16have been shown to play crucial roles in the development of multidrug resistance in gastric cancer cells.(26)In our case-control study, we found that the variant genotypes of rs895819located at hsa-mir-27a conferred a48%increased risk of developing gastric cancer in the Chinese population.Hu et al.also indicated that SNP(rs11614913)in hsa-mir-196a2and variant genotypes (rs3746444)in hsa-mir-499were associated with significantly increased risks of breast cancer in Chinese women.Moreover, they suggested that a variant homozygote of rs11614913in hsa-mir-196a2had poor survival in patients with non-small cell lung cancer.(18,27)Therefore,it has been indicated that polymorphism within miRNA precursors might be used as candidate biomar-kers for cancer susceptibility.In addition,our stratified analyses revealed that the functional polymorphism was associated with the increased risk of gastric cancer among subgroups of older subjects(age>58years),but not among younger subjects.Carcinogenesis is considered as accumulation of genetic events during aging,and gastric cancer has a steep slope for the age-specific increase in incidence.(28) The increased risk that was observed in older subjects implies that the gene–environment interaction might be involved in car-cinogenesis and the hsa-mir-27a genotype effects tended to be age specific.We also found that an elevated risk was evident in rural subjects.Interestingly,we unexpectedly found that male subjects with the variant genotypes were associated with a53% increased risk of gastric cancer.Previously,we also noted the gender-specific associations between the polymorphism and phenotypes.(20)Arisawa et al.(29)also found that the miR-27a polymorphism was associated with the gastric mucosal atrophy only in male subjects.Further studies are needed to clarify the underlying mechanism.More importantly,a significant associa-tion of hsa-mir-27a variant genotypes with lymph node metasta-sis was observed.A recent study also found a correlation between miR-27a and lymph node metastasis in undifferentiated gastric cancer.(30)It suggested that miR-27a,as one of the poten-tial oncomirs,might contribute to tumor invasion and meta-stasis.Recent studies have demonstrated that mutation or SNP in miRNAs could influence miRNAs expression and function.(31,32) Zeng et al.(33)have shown that the integrity and secondary structure of pre-miRNAs was crucial to the processing and mat-uration of miRNAs.One particular SNP located at the seed region of mature miR-125a could significantly affect pre-miRNA maturation and reduce miRNA-mediated posttranscrip-tional suppression.(9)Consistently,Jazdzewski et al.(24)indi-cated that a common polymorphism in pre-miR-146a could contribute to genetic predisposition to papillary thyroid carci-noma through affecting the expression of miR-146.This increas-ing evidence encouraged us to investigate the possible role of this SNP rs895819in regulating expression of miR-27a and its target genes.On the basis of the model of Zeng and Cullen, mutations that took place in the stem of pre-miRNAs had a marked effect on miRNAs processing.(33)Although the poly-morphism within miR-27a was observed in the terminal loop, our results also revealed that there was a significant influence on miR-27a expression based on genotypes.One possible reason is that a mutant with terminal loop may slightly impair processing of pri-miRNA and DGCR8binding based on the ssRNA-dsRNA Junction Anchoring Model.(34)We are thefirst tofind that functional polymorphism might be responsible for elevated miR-27a expression levels,which were associated with significantly reduced ZBTB10mRNA.Thus,we95.00P < 0.001P = 0.003P = 0.002r = –0.37990.00miRNA27a/U6(1%)ZBTB1/TBP(1%)85.0080.0075.0065.00130.00120.00110.00100.0090.0080.00ZBTB1/TBP(1%)130.00120.00110.00100.0090.0080.0065.0070.0075.0080.00miRNA27a/U6 (100%)85.0095.0090.00AA(n = 25)AG(n = 30)GG(n = 10)AA(n = 25)AG(n = 30)GG(n = 10)70.00(A)(B)(C)Fig.2.Relationship between rs895819genotypes and expression of miR-27a or Zincfinger and BTB domain containing10(ZBTB10).(A) Expression of miRNA-27a was determined by TaqMan MicroRNA Assays(Applied Biosystems Inc.).Normalization was performed with RNU6B.The box plot revealed the distribution of the miR-27a levels within each genotype class in65gastric cancer cases.(B)The relative expression of ZBTB10mRNA normalization was performed with TATA binding protein(TBP)and determined by SYBR Green real-time PCR. The box plot revealed the distribution of the ZBTB10mRNA levels within each genotype class in65gastric cancer cases.ANOVA with Dunnett post-test,compared with the AA group.(C)Correlation between mRNA ZBTB10and expression of miR-27a in65gastric cancer cases(r=)0.379;P=0.002).Sun et al.Cancer Sci|October2010|vol.101|no.10|2245investigated whether ZBTB10mRNA was negatively regulated by miR-27a in gastric cancer.As shown in Fig.2C,ZBTB10lev-els inversely correlated with miR-27a levels in patients with gas-tric cancer.This suggests ZBTB10as a predominant target of miR-27a.ZBTB10,a novel zincfinger protein,could regulate gastrin gene expression by interfering with Sp1transactiva-tion.(15)There is growing evidence that Sp1–Sp4proteins play a critical role in the growth and metastasis of many tumors by reg-ulating expression of multiple genes such as VEGF,VEGFR1, VEGFR2and survivin.(35,36)In particular,Sp1protein is widely overexpressed in various human tumors including breast carci-noma,colorectal cancer and gastric carcinoma.(37)Moreover, strong Sp1expression was more frequently observed at an advanced disease stage among gastric carcinoma patients.(38) Therefore,combined with previous observations,it is biologi-cally plausible that miR-27a,through regulating ZBTB10,results in overexpression of Sp proteins and Sp-dependent genes,which play important roles in gastric cancer cell survival and angiogen-esis.(13,38)Potential limitations of the current study should be consid-ered.First,the design was a hospital-based case-control study, and the controls were recruited from patients with non-malig-nant diseases.This could lead to the possibility of selection bias. Nonetheless,the genotype distribution of the controls was in Hardy–Weinberg equilibrium.Second,in the present study,only mature miR-27a expression was examined;whether the miR-27a SNP might also influence the processing of pri-miRNA and⁄or pre-miRNA still needs to be verified.Third,the sample size of the present study was relatively small,particularly for stratified analyses,for evaluating the relationship between the amount of miR-27a and tumor invasion or metastasis.Finally,Helicobacter pylori infection is one of the independent risk fac-tors of gastric cancer.We did not have enough information on Helicobacter pylori status,because it was unethical to do Heli-cobacter pylori tests in every subject,especially in the controls. Therefore,further studies with a larger sample size are warranted.However,our observations provided valuable insights and interesting information and might serve to guide future studies in this area.In conclusion,we are thefirst to provide evidence that genetic variants in hsa-mir-27a conferred not only an increased risk of gastric cancer but also with metastasis of gastric cancer in the Chinese population,possibly by increasing the levels of miR-27a,which is accompanied by reduced ZBTB10mRNA. We expect ourfindings will provide novel insights into the molecular mechanisms of tumorigenesis and could eventually contribute to the development of early diagnosis for gastric cancer.AcknowledgmentsThis work was supported by the following grants:National Natural Sci-ence Foundation of China(No.30873099);Natural Science Foundation of Jiangsu Province(No.BK2006525);Natural Science Foundation of Jiangsu Provincial Education Office(No.08KJB320004, KY101J28019);‘‘333Project’’,‘‘Qinglan Project’’and‘‘Six RenCai GaoFeng Project’’Funding for the Young Academic Leader of Jiangsu Province.Disclosure StatementThe authors have no conflict of interest.References1Parkin DM,Bray F,Ferlay J,Pisani P.Global cancer statistics,2002.CA Cancer J Clin2005;55:74–108.2Sun J,Misumi J,Shimaoka A,Aoki K,Esaki F.Stomach cancer-related mortality.Eur J Cancer Prev2001;10:61–7.3Bartel DP.MicroRNAs:genomics,biogenesis,mechanism,and function.Cell 2004;116:281–97.4Chan JA,Krichevsky AM,Kosik KS.MicroRNA-21is an 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miR-27a对两种结肠癌细胞增殖和侵袭能力的影响张子雯;傅小一【摘要】目的探讨microRNA-27a(miR-27a)对2种结肠癌细胞增殖和侵袭能力的影响.方法应用miR-27a反义寡核苷酸(antisense oligonucleotides,ASO)在体外转染结肠癌细胞系SW620和HCT116,设置转染无义序列组作为对照组.RT-PCR 检测细胞miR-27a表达;MTT法检测细胞增殖;Transwell小室侵袭实验检测细胞侵袭能力;Western blot检测转染细胞FOXO1蛋白表达.结果与对照组相比,转染miR-27a ASO后结肠癌细胞系SW620和HCT116的miR-27a表达水平均明显降低(P＜0.001),增殖能力和侵袭能力均受到明显抑制(P＜0.01),FOXO1蛋白表达均明显增高(P＜0.05).结论抑制miR-27a表达可显著降低结肠癌细胞系SW620和HCT116的增殖能力和侵袭能力,而调控下游FOXO1蛋白表达可能是miR-27a 参与结肠癌细胞增殖和侵袭的主要机制.【期刊名称】《中国生化药物杂志》【年(卷),期】2015(000)002【总页数】4页(P24-27)【关键词】miR-27a;结肠癌;增殖;侵袭;FOXO1【作者】张子雯;傅小一【作者单位】江西省宜春学院医学院,江西宜春336000;江西省宜春学院医学院,江西宜春336000【正文语种】中文【中图分类】R735.3结肠癌是常见的消化道恶性肿瘤之一。

近年来，随着人们生活方式及饮食习惯的改变，国内外结肠癌发病率及死亡率均呈逐年上升趋势，严重威胁患者的生命健康[1]。

但目前对于结肠癌增殖及侵袭的机制尚未完全明确。

非蛋白质编码微小RNA(microRNA， miRNA)的出现为恶性肿瘤的研究提供了新思路[2]。

越来越多证据显示，miRNA与肿瘤密切相关，miRNA可通过与靶mRNA互补配对，从而调控蛋白翻译过程，在肿瘤细胞增殖、分化、凋亡和基因调控中发挥重要作用[3]。

超微血管成像技术诊断乳腺肿瘤的meta分析陈思晗;熊虎;税典雅;高小瞻;刘泽伟【期刊名称】《临床荟萃》【年(卷),期】2024(39)2【摘要】目的采用meta分析方法评价超微血管成像技术(SMI)评估乳腺肿瘤良恶性的临床价值。

方法系统搜索PubMed、Cochrane Library、Embase、CNKI、CBM、WANFANG、VIP中建库至2022年9月20日有关超微血流成像(super microvascular imaging,SMI)评估乳腺肿瘤良恶性的文献。

由2名研究人员依据纳入及排除标准初筛文献并提取信息;运用QUADAS-2工具评估纳入原始文献质量,Review Manager 5.3绘制文献质量评估图,Stata 17.0计算SMI评估乳腺肿瘤的敏感度(SEN)、特异度(SPE)、阳性似然比(PLR)、阴性似然比(NLR)及诊断比值比(DOR),绘制综合受试者工作特征曲线(SROC),获得曲线下面积;绘制漏斗图进行发表偏倚评估。

结果最终纳入15篇文献、1769例乳腺肿瘤患者、共1912个病灶,其中恶性结节916个、良性结节991个。

Meta分析结果显示,SMI评估乳腺病变SEN、SPE、PLR、NLR、DOR及SROC曲线下面积分别为0.82[95%CI(0.79~0.84)]、0.87[95%CI(0.85~0.89)]、6.58[95%CI(4.58~9.43)]、0.19[95%CI(0.16~0.24)]、39.47[95%CI(27.18~57.31)]、0.93[95%CI(0.90~0.95)]。

结论SMI评估乳腺良恶性病变的鉴别具有较高的诊断准确性。

【总页数】7页(P108-114)【作者】陈思晗;熊虎;税典雅;高小瞻;刘泽伟【作者单位】宜昌市第二人民医院超声科【正文语种】中文【中图分类】R737.9【相关文献】1.超微血管成像技术诊断乳腺肿瘤的临床研究进展2.超微血管成像技术对乳腺纤维腺瘤及乳腺叶状肿瘤的鉴别诊断价值3.超微血管成像技术对BI-RADS 4类乳腺微小肿瘤的诊断价值4.超微血管成像技术在乳腺肿瘤诊断中的应用进展因版权原因，仅展示原文概要，查看原文内容请购买。

磁共振动态增强参数及miR-27、miR-155表达与乳腺癌的关联性秦恺;叶飞;顾桂源;贲腾;陆松华【期刊名称】《影像科学与光化学》【年(卷),期】2022(40)2【摘要】探究磁共振动态增强、miR-27、miR-155表达与乳腺癌的关联性。

选取100例乳腺癌患者作为观察组,另选取60例乳腺良性病变患者作为对照组。

比较两组DCE-MRI定量参数[转运常数(K_(trans))、速率常数(K_(ep))、血管外细胞间隙体积分数(c)]与miR-27、miR-155表达,进行相关性分析,并评价其诊断效能。

结果显示,观察组K_(trans)、K_(ep)、miR-27和miR-155表达水平较对照组高(P<0.05);K_(trans)、K_(ep)与miR-27和miR-155表达水平呈显著正相关(P<0.05);K_(trans)、K_(ep)、miR-27、miR-155为乳腺癌发生的独立影响因素(P<0.05);K_(trans)、K_(ep)、miR-27和miR-155表达与组织学分级、临床分期、淋巴结转移、孕激素及雌激素受体表达有关(P<0.05);DCE-MRI定量参数、miR-27和miR-155表达联合诊断乳腺癌的AUC为0.905。

乳腺癌发生DCE-MRI定量参数(K_(trans)、K_(ep))与miR-27、miR-155表达水平均上调,且与病理特征有关。

【总页数】6页(P263-268)【作者】秦恺;叶飞;顾桂源;贲腾;陆松华【作者单位】南通大学附属海安医院胸外科【正文语种】中文【中图分类】R73【相关文献】1.动态增强磁共振成像定量参数评估乳腺癌新辅助化疗效果及相关性2.动态增强磁共振成像定量参数对乳腺癌新辅助化疗疗效评价及相关性研究3.动态增强磁共振成像定量参数早期预测局部进展期乳腺癌新辅助化疗效果的价值4.动态增强磁共振成像定量参数早期预测局部进展期乳腺癌新辅助化疗效果的价值5.动态增强磁共振成像定量参数对乳腺癌新辅助化疗疗效评价及相关性研究因版权原因，仅展示原文概要，查看原文内容请购买。