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长链编码RNA GSTM3TV2作为一种癌基因促进胰腺癌的化疗耐药

[The long coding RNA GSTM3TV2 acts an oncogene to promote chemoresistance in pancreatic cancer].

作者信息

Xiong G B, Yang G, Feng M Y, Zhao F Y, Zheng L F, Zhang T P, Zhao Y P

机构信息

Department of General Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, China.

Department of Nuclear Medicine, Peking Union Medical College Hospital, Peking Union Medical College, Academy of Chinese Medical Sciences, Beijing 100730, China.

出版信息

Zhonghua Wai Ke Za Zhi. 2019 Sep 1;57(9):691-697. doi: 10.3760/cma.j.issn.0529-5815.2019.09.009.

Abstract

To examine the expression of the long coding RNA GSTM3TV2 in pancreatic cancer tissues and to examine its role and mechanism in chemoresistance of pancreatic cancer cells. The expression of lncRNA GSTM3TV2 in 15 pancreatic cancer specimens and corresponding adjacent to cancer tissue samples diagnosed by Department of Pathology, Peking Union Medical College Hospital was detected by real-time PCR.And the expressions of GSTM3TV2 in pancreatic cancer cell AsPC-1, BxPC-3, MIAPaCa-2, PanC-1, SU86.86, T3M4, and chemoresistant cells AsPC-1/GR and MIAPaCa-2/GR, and human pancreatic nestin-expressing cells hTERT-HPNE were detected. Pancreatic cancer cell lines were transfected with GSTM3TV2-pcDNA3.1(+)in order to get cells with GSTM3TV2 overexpression.GSTM3TV2-siRNA was transfected into pancreatic cancer cells to knock down GSTM3TV2. The cell chemoresistance was measured by CCK-8 and flow cytometry assay when incubated with nab-paclitaxel. At the same time, subcutaneous xenograft tumor models were established in nude mice to observe the effect of GSTM3TV2 on chemoresistance of tumor growth in nude mice.Western blot assay was also performed to detect the molecular mechanism of chemoresistance of GSTM3TV2. Comparing toadjacent tissues(0.084±0.019), GSTM3TV2 expression was significantly upregulated in the pancreatic cancer tissues(0.493±0.084) (5.146, 0.05). GSTM3TV2 expression were higher in the chemotherapy resistance pancreatic cancer cells AsPC-1/GR(210.799±19.788) and MIAPaCa-2/GR(122.408±23.419) than that in the AsPC-1(3.793±0.615) and the MIAPaCa-2(5.179±1.095)(21.800,0.05;-18.490,0.05). The results of in vivo experiments showed that the volume of subcutaneously transplanted tumors in the overexpressing GSTM3TV2 group ((1 059.609±102.498)mm(3)) was significantly larger than that in the control group((566.414±81.087) mm(3)) by treated with nab-paclitaxel(4.230,0.05).Meanwhile, GSTM3TV2 could promote the expression of Cyclin D1, CDK6, Cyclin E1, Vimentin, N-cadherin, ZEB1, Snail and Slug; but decrease cleaved caspase-3, cleaved PARP in pancreatic cancer cells. The expression level of GSTM3TV2 in pancreatic canceris higher than that in paired adjacent tissues. GSTM3TV2 may act as an oncogene to promote chemoresistance in pancreatic cancer through regulation of cell proliferation, apoptosis, and epithelial-mesenchymal transition.

摘要

检测长链编码RNA GSTM3TV2在胰腺癌组织中的表达,并探讨其在胰腺癌细胞化疗耐药中的作用及机制。采用实时荧光定量PCR检测北京协和医院病理科诊断的15例胰腺癌标本及相应癌旁组织中lncRNA GSTM3TV2的表达。检测GSTM3TV2在胰腺癌细胞AsPC-1、BxPC-3、MIAPaCa-2、PanC-1、SU86.86、T3M4及化疗耐药细胞AsPC-1/GR和MIAPaCa-2/GR以及人胰腺巢蛋白表达细胞hTERT-HPNE中的表达。将GSTM3TV2-pcDNA3.1(+)转染至胰腺癌细胞系,以获得GSTM3TV2过表达细胞。将GSTM3TV2-siRNA转染至胰腺癌细胞以敲低GSTM3TV2。用CCK-8法和流式细胞术检测细胞与纳米白蛋白紫杉醇孵育后的化疗耐药性。同时,建立裸鼠皮下移植瘤模型,观察GSTM3TV2对裸鼠肿瘤生长化疗耐药的影响。采用蛋白质免疫印迹法检测GSTM3TV2化疗耐药的分子机制。与癌旁组织(0.084±0.019)相比,GSTM3TV2在胰腺癌组织中的表达显著上调(0.493±0.084)(5.146,P<0.05)。GSTM3TV2在化疗耐药的胰腺癌细胞AsPC-1/GR(210.799±19.788)和MIAPaCa-2/GR(122.408±23.419)中的表达高于AsPC-1(3.793±0.615)和MIAPaCa-2(5.179±1.095)(21.800,P<0.05;-18.490,P<0.05)。体内实验结果显示,经纳米白蛋白紫杉醇处理后,GSTM3TV2过表达组皮下移植瘤体积((1 059.609±102.498)mm³)明显大于对照组((566.414±81.087)mm³)(4.230,P<0.05)。同时,GSTM3TV2可促进胰腺癌细胞中细胞周期蛋白D1、细胞周期蛋白依赖性激酶6、细胞周期蛋白E1、波形蛋白、N-钙黏蛋白、锌指蛋白E盒结合因子1、蜗牛蛋白和蛞蝓蛋白的表达;但可降低胰腺癌细胞中裂解的半胱天冬酶-3、裂解的聚(ADP-核糖)聚合酶的表达。胰腺癌中GSTM3TV2的表达水平高于配对的癌旁组织。GSTM3TV2可能作为癌基因,通过调节细胞增殖、凋亡和上皮-间质转化促进胰腺癌的化疗耐药。

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