RNAi抑制PSA表达对前列腺癌22RV1细胞增殖和侵袭力的影响

目的 研究PSA表达对去势抵抗性前列腺癌22RV1细胞增殖和侵袭的影响.方法 构建含有针对PSA基因特异性短发夹RNA(shRNA,分别为shPSA1、shPSA2、shPSA3、shPSA4)和阴性对照序列(NC)的慢病毒载体并由慢病毒颗粒包装后,分别感染去势抵抗性前列腺癌22RV1细胞并筛选稳定表达PSA特异性shRNA的shPSA1-、shPSA2-、shPSA3-、shPSA4-和表达NC序列的NC-22RV1细胞.分别应用实时荧光定量聚合酶链反应和Western blot检测shRNA对22RV1细胞中PSA mRNA及蛋白表达的影响;采用CCK-8法、Transwell侵袭试验和流式细胞术检测PSA基因沉默后的22RV1细胞增殖、侵袭和凋亡情况.结果 与NC-22RV1细胞相比,shPSA3-22RV1细胞的基因和蛋白表达水平的沉默效果最好;shPSA2组在24、48、72 h的吸光度值分别为(0.1564±0.0225)、(0.2438±0.0358)、(0.3641±0.0205),shPSA3组分别为(0.1069±0.0120)、(0.1588±0.0192)、(0.2534±0.0289),与NC组比较差异均有统计学意义(P<0.01);shPSA1、shPSA2、shPSA3组侵袭力分别为NC组的77.35%(P=0.002)、54.35%(P<0.001)、42.21%(P<0.001);shPSA2、shPSA3组凋亡率分别为(27.97±4.28)%(P=0.001)、(43.03±3.19)%(P<0.001),高于NC组[(16.77±0.59)%].结论 PSA具有促进去势抵抗性前列腺癌22RV1细胞的增殖和侵袭效应,抑制PSA表达后前列腺癌22RV1细胞凋亡增加.

Effects of PSA expression inhibition by RNA interference on the cancer cells proliferation and invasion in human prostate cancer 22RV1 cells

Objective To study the effects of prostate specific antigen (PSA)on the prolifera-tion and invasion of human prostatic cancer 22RV1 cells. Methods PSA-targeted shRNA lentivirus vectors containing shPSA1,shPSA2,shPSA3,shPSA4 or negative control (NC)were developed, and then were packaged in lentiviral particles,which were then used to infect castration-resistant prostatic cancer 22RV1 cells for the generation of shPSA1-,shPSA2-,shPSA3-,shPSA4-,or NC-22RV1 cells respectively.The expression levels of PSA mRNA and protein were detected by real-time fluorescent quantitative-PCR (qRT-PCR)and western blot,respectively.The proliferation,in-vasion abilities and apoptosis of 22RV1 cells were evaluated by CCK-8 assay,transwell cell invasion assay and flow cytometry respectively. Results The expression levels of PSA mRNA were down-regulated in shPSA1-22RV1,shPSA2-22RV1,and especially shPSA3-22RV1 cells which showed the most significantly gene silencing efficacy (P <0.01).The expression levels of protein was significant-ly lower in shPSA3-22RV1 cells than in NC-22RV1 cells (P <0.01 ).The proliferation abilities of shPSA2-and shPSA3-22RV1 cells at 24,48 and 72 h after adherence were obviously inhibited as compared with those of NC-22RV1 cells (all P <0.01 ).The invasion ability of shPSA1-,shPSA2-and shPSA3-22RV1 cells were all decreased (P <0.01).Cell apoptosis analysis by cell cytometry showed that the transfection of shPSA2 and shPSA3 construct increased the proportion of apoptotic cells (P <0.01). Conclusions PSA promotes CRPC 22RV1 cell proliferation,invasion,and inhibites its apoptosis.