组织因子途径抑制物-2抑制人前列腺癌细胞的上皮间质转化  

Tissuefactor pathway inhibitor-2 inhibits epithelial-mesenchymal transition in human prostate cancer cells

作　　者：刘树瀚 宣强[1] 谈宜傲[1] 吴本鹤 孙凌峰[1] 乔龙标 翟路路 孙友文[1] 周林玉[1] 

机构地区：[1]安徽医科大学附属省立医院泌尿外科,合肥230001 [2]安徽医科大学附属省立医院胆胰外科,合肥230001

出　　处：《安徽医科大学学报》2015年第12期1738-1743,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:1408085MH181)

摘　　要：目的探讨组织因子途径抑制物-2(TFPI-2)对人前列腺癌细胞上皮间质转化的影响。方法将人前列腺癌细胞株PC3M进行培养,并分为3组,分别转染TFPI-2基因真核表达载体、空载体及不转染,细胞划痕实验和细胞侵袭小室实验检测3组细胞的迁移能力和侵袭能力,用转化生长因子-β1(TGF-β1)处理细胞,倒置相差显微镜下观察3组细胞形态,Western blot法和逆转录聚合酶链式反应(RT-PCR)法分别检测转染组、转染空载体组及未转染组上皮间质转化标志物-E钙黏素(E-cadherin)、波形蛋白(vimentin)和snail在蛋白和mRNA水平上的表达量。结果细胞划痕实验和细胞侵袭小室实验结果显示,转染TFPI-2基因真核表达载体的PC3M细胞迁移率较低(P<0.05),侵袭细胞数较少(P<0.05);显微镜下观察显示转染组细胞大多呈上皮型,转染空载体组及未转染组细胞大多呈间叶型;Western blot法和RT-PCR法检测结果显示,无论在蛋白水平还是mRNA水平,转染组较转染空载体组及未转染组E-cadherin表达产物较多,vimentin和snail蛋白表达产物较少,差异有统计学意义(P<0.05),转染空载体组及未转染组差异无统计学意义。结论 TFPI-2可以抑制人前列腺癌细胞的上皮间质转化,这是其抑制前列腺癌细胞侵袭与转移的可能机制之一,为基因及生物靶向治疗前列腺癌提供新的思路和方向。Objective To investigate the influence of tissue factor pathway inhibitor-2 （TFPI-2） on epithelial-mes- enchymal transition （EMT） in human prostate cancer ceils. Methods PC3M cells were cultured and divided into three groups, respectively transfected with TFPI-2 gene eukaryotic expression vector, empty carrier and nothing. Wound healing assay and thanswell assay were performed to evaluate motility and invasive capacity of three groups PC3M cells. Then the three groups ceils were dealt with by transforming growth factord31 （ TGF-131 ）, cellular mor- phology was observed with inverted phase contrast microscope. Protein and mRNA levels of characteristic markers （E-cadherin, vimentin and snail） involved in EMT were respectively detected in cells of three groups by Western blot and reverse transcription polymerase chain reaction （RT-PCR）. Results were statistically analyzed. Results Wound healing assay and thanswell assay revealed that migration rate of transfected cells was lower （ P 〈 0.05 ）, in- vasive cells number of that was fewer （P 〈 0. 05）. Transfected cells displayed an epithelial-like morphology, trans- fected empty-carrier ceils and non-transfected cells showed a mesenchymal-like morphology by inverted phase con- trast microscope. Western blot and RT-PCR displayed that E-cadherin was more, vimentin and snail were less on protein and mRNA levels in transfected ceils compared with transfected empty-carrier cells and non-transfected cells; the difference was statistically significant （P 〈 0. 05 ）, and the difference between transfected empty-carrier cells and non-transfected cells was not statistically significant . Conclusion TFPI-2 can inhibit EMT in human prostate cancer ceils. It is one of the possible mechanisms of inhibiting the invasion and metastasis of prostate cancer cell. It will help to provide new train of thought and direction for gene and biologically targeted theropy for pros- tate cancer.

关 键 词：组织因子途径抑制物-2 前列腺癌 上皮间质转化 

分 类 号：R737.25[医药卫生—肿瘤]