miRNA-30c及其靶基因MTA-1在子宫内膜癌裸鼠皮下移植瘤中的作用
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摘要
目的:探讨过表达miRNA-30c对子宫内膜癌裸鼠皮下移植瘤生长的影响。方法:应用子宫内膜癌Ishikawa细胞系建立子宫内膜癌裸鼠皮下移植瘤模型,予裸鼠皮下移植瘤内多点注射miRNA-30c前体表达质粒,并以阴性质粒及PBS为对照。测量移植瘤体积,计算肿瘤生长抑制率;荧光定量PCR技术检测移植瘤组织miRNA-30c表达水平;蛋白质印迹技术检测移植瘤组织MTA-1蛋白表达水平;免疫组化技术检测移植瘤组织E-cadherin表达及微血管密度(MVD)。结果:成功建立子宫内膜癌裸鼠皮下移植瘤模型,miRNA-30c质粒治疗明显抑制肿瘤生长,抑制率为(46.98±5.13)%(P<0.01);上调移植瘤组织中miRNA-30c的表达(P<0.05),下调MTA-1蛋白表达(P<0.05),下调移植瘤组织MVD(P<0.05),上调移植瘤组织E-cadherin表达(P<0.05)。结论:过表达miRNA-30c可以通过下调MTA-1表达明显抑制子宫内膜癌裸鼠皮下移植瘤的生长。
Objective: To investigate the inhibition effect on tumor growth by up-regulating the expression of miRNA-30 c in nude mice model. Methods: Subcutaneous xenografts of mude mice models with Ishikawa cells were established,the mice were treated with miRNA-30 c expression plasmid intratumorally,controled by negative control plasmid and PBS. Then, the xenograft volume was measured and inhibition ratio of tumor proliferation was calculated. Moreover,the expression of miRNA-30 c was analysed by Realtime-PCR and the expression of MTA-1protein was analysed by Western blot,the microvessel density and expression of E-cadherin were detected by immunohistochemistry. Results: The subcutaneous xenografts models of mude mice were established successfully.Treatment with miRNA-30 c expression plasmid inhibited the growth of xenograft and the inhibitory rate was( 46. 98 ±5. 13) %( P < 0. 01). The expression of miRNA-30 c was up-regulated( P < 0. 05) and MTA-1 protein expression was down-regulated( P < 0. 05) respectively. Immunohistochemistry showed that the microvessel density decreased( P < 0. 05),the expression of E-cadherin increased( P < 0. 05). Conclusion: Up-regulation of miRNA-30 c can inhibit the growth of xenograft in nude mice model of endometrial carcinoma by down-regulating MTA-1.
Objective: To investigate the inhibition effect on tumor growth by up-regulating the expression of miRNA-30 c in nude mice model. Methods: Subcutaneous xenografts of mude mice models with Ishikawa cells were established,the mice were treated with miRNA-30 c expression plasmid intratumorally,controled by negative control plasmid and PBS. Then, the xenograft volume was measured and inhibition ratio of tumor proliferation was calculated. Moreover,the expression of miRNA-30 c was analysed by Realtime-PCR and the expression of MTA-1protein was analysed by Western blot,the microvessel density and expression of E-cadherin were detected by immunohistochemistry. Results: The subcutaneous xenografts models of mude mice were established successfully.Treatment with miRNA-30 c expression plasmid inhibited the growth of xenograft and the inhibitory rate was( 46. 98 ±5. 13) %( P < 0. 01). The expression of miRNA-30 c was up-regulated( P < 0. 05) and MTA-1 protein expression was down-regulated( P < 0. 05) respectively. Immunohistochemistry showed that the microvessel density decreased( P < 0. 05),the expression of E-cadherin increased( P < 0. 05). Conclusion: Up-regulation of miRNA-30 c can inhibit the growth of xenograft in nude mice model of endometrial carcinoma by down-regulating MTA-1.
引文
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[24]池丽敏,肖鹏,陈定科,等.Vimentin蛋白在子宫内膜样腺癌组织中的表达及其临床意义[J].现代医学,2015,43(10):1216-1218.
[2]LAM E W,SHAH K,BROSENS J J.The diversity of sex steroid action:the role of micro-RNAs and FOXO transcription factors in cycling endometrium and cancer[J].J Endocrinol,2012,212(1):13-25.
[3]JEMAL A,BRAY F,CENTER M M,et al.Global cancer statistics[J].CA Cancer J Clin,2011,61(2):69-90.
[4]ZHOU H J,XU X F,XUN Q Y,et al.microRNA-30c negatively regulates endometrial cancer cells by targeting metastasisassociated gene-1[J].Oncol Rep,2012,27(3):807-812.
[5]KONG X Y,XU X F,YAN Y H,et al.Estrogen regulates the tumour suppressor MiRNA-30c and its target gene,MTA-1,in endometrial cancer[J].PLo S One,2014,9:90810.
[6]YANOKURA M,BANNO K,IIDA M,et al.MicroRNAS in endometrial cancer:recent advances and potential clinical applications[J].Excli J,2015,14:190-198.
[7]LI S,YANG C,ZHAI L,et al.Deep sequencing reveals small RNA characterization of invasive micropapillary carcinomas of the breast[J].Breast Cancer Res Treat,2012,136(1):77-87.
[8]HUANG Z,ZHANG L,YI X,et al.Diagnostic and prognostic values of tissue hsa-miR-30c and hsa-miR-203 in prostate carcinoma[J/OL].T Tumour Biol,2015,doi:10.1007/s13277-015-4262-9.
[9]SAISELET M,GACQUER D,SPINETTE A,et al.New global analysis of the microRNA transcriptome of primary tumors and lymph node metastases of papillary thyroid cancer[J].BMC Genomics,2015,16:828.
[10]ZHANG Q,YU L,QIN D,et al.Role of microRNA-30c targeting ADAM19 in colorectal cancer[J].PLo S One,2015,10(3):e0120698.
[11]LIU D,WU J,LIU M,et al.Downregulation of miRNA-30c and miR-203a is associated with hepatitis C virus core protein-induced epithelial-mesenchymal transition in normal hepatocytes and hepatocellular carcinoma cells[J].Biochem Biophys Res Commun,2015,464(4):1215-1221.
[12]RODRIGUEZ-GONZALEZ F G,SIEUWERTS A M,SMID M,et al.MicroRNA-30c expression level is an independent predictor of clinical benefit of endocrine therapy in advanced estrogen receptor positive breast cancer[J].Breast Cancer Res Treat,2011,127(1):43-51.
[13]EGELAND N G,LUNDES S,JONSDOTTIR K,et al.The role of MicroRNAs as predictors of response to tamoxifen treatment in breast cancer patients[J].Int J Mol Sci,2015,16(10):24243-24275.
[14]HEINZELMANN J,HENNING B,SANJMYATAV J,et al.Specific miRNA signatures are associated with metastasis and poor prognosis in clear cell renal cell carcinoma[J].World J Urol,2011,29(3):367-373.
[15]NGUYEN H T,JIA W,BEEDLE A M,et al.Lysophosphatidic acid mediates activating transcription factor 3 expression which is a target for post-transcriptional silencing by miR-30c-2-3p[J].PLo S One,2015,10(9):e0139489.
[16]WU W,ZHANG X,LIAO Y,et al.miR-30c negatively regulates the migration and invasion by targeting the immediate early response protein 2 in SMMC-7721 and Hep G2 cells[J].Am J Cancer Res,2015,5(4):1435-1446.
[17]BUSACCA S,GERMANO S,DE L,et al.MicroRNA signature of malignant mesothelioma with potential diagnostic and prognostic implications[J].Am J Respir Cell Mol Biol,2010,42(3):312-319.
[18]LEE H,PARK C S,DEFTEREOS G,et al.MicroRNA expression in ovarian carcinoma and its correlation with clinicopathological features[J].World J Surg Oncol,2012,10:174.
[19]GAROFALE M,ROMANO G,DI G,et al.EGFR and MET receptor tyrosine kinase-altered microRNA expression induces tumorigenesis and gefitinib resistance in lung cancers[J].Nat Med,2012,18(1):74-82.
[20]BALASENTHIL S,BROADDUS R R,KUMAR R.Expression of metastasis-associated protein 1(MTA1)in benign endometrium and endometrial adenocarcinomas[J].Hum Pathol,2006,37(6):656-661.
[21]NAGARAJ S R,SHILPA P,RACHAIAH K,et al.Crosstalk between VEGF and MTA1 signaling pathways contribute to aggressiveness of breast carcinoma[J].Mol Carcinogen,2015,54(5):333-350.
[22]TUNCAY C S,CIMEN I,SAVAS B,et al.MTA-1 expression is associated with metastasis and epithelial to mesenchymal transition in colorectal cancer cells[J].Tumour Biology,2013,34(2):1189-1204.
[23]WENG W,YIN J,ZHANG Y,et al.Metastasis-associated protein 1 promotes tumor invasion by downregulation of Ecadherin[J].Int J Oncol,2014,44(3):812-818.
[24]池丽敏,肖鹏,陈定科,等.Vimentin蛋白在子宫内膜样腺癌组织中的表达及其临床意义[J].现代医学,2015,43(10):1216-1218.