Adv-shRNA抑制CXCR4基因表达对人前列腺癌细胞PC-3增殖和侵袭力的影响  

Effect of CXCR4 expression inhibited by Adv-shRNA on the proliferation and invasiveness of PC-3 cells in prostate carcinoma

作　　者：李自智[1] 佟明[1] 黄伟超[1] 金艳阳[1] 

机构地区：[1]辽宁医学院第一附属医院,辽宁锦州121001

出　　处：《现代预防医学》2013年第12期2316-2319,2322,共5页Modern Preventive Medicine

摘　　要：目的通过特异性抑制前列腺癌细胞株PC-3中趋化因子受体4(CXCR4)基因的表达,检测RNA干扰片段对PC-3细胞增殖及侵袭转移能力的影响。方法将CXCR4的特异性RNA干扰腺病毒载体转染至前列腺癌PC-3细胞72h后荧光显微镜观察转染情况;采用RT-PCR法检测转染后CXCR4基因mRNA表达情况;Western blot检测转染后CXCR4蛋白表达情况;MTT法检测PC-3细胞增殖情况;Transwell小室侵袭实验检测对PC-3细胞侵袭转移能力。结果 (1)成功将CXCR4-shRNA重组腺病毒载体转染至前列腺癌细胞PC-3;(2)重组腺病毒转染组CXCR4基因mRNA及蛋白均低于阴性对照组和空白组,差异有统计学意义(P﹤0.01),而阴性对照组与空白组之间差异无统计学意义(P﹥0.05);(3)阴性对照组与空白组细胞增殖迅速,两组之间差异无统计学意义(P﹥0.05),实验组经腺病毒载体转染后细胞增殖程度显著减少,与前两组相比差异有统计学意义(P﹤0.05);(4)CXCR4-shRNA腺病毒载体与空白组及对照组相比能显著抑制PC-3细胞的侵袭力(P﹤0.05),抑制率为57.01%,空白组与对照组相比差异无统计学意义(P﹥0.05)。结论重组腺病毒载体CXCR4-shRNA可有效抑制前列腺癌细胞PC-3中CXCR4基因的mRNA和蛋白的表达,并在一定程度上抑制癌细胞增殖及远处侵袭转移,可作为动物实验的理论基础和前期条件。OBJECTIVE To detect the effect of RNA interference fragment on the ability of proliferation and invasiveness of PC-3 cells in prostate carcinmna with specific inhibition of CXCR4 gene expression. METHODS CXCR4 adenovirus vector interfered by RNA was transferred into PC-3 cells and the transfer situation was observed after 72 hours. The expression of mRNA in CXCR4 gene was detected by RT-PCR, the proliferation of PC-3 cells in prostate carcinoma was detected by MTT and the protein expression of CXCR4 after transferation was detected by Western blot. The invasive ability of PC-3 cells was detected by Transwell cabinet invasion experiment. RESULTS （ 1 ） The transferation of CXCR4-shRNA recombinant aden-ovirns vector into PC-3 cells of prostate carcinoma was successful; （2） The level of CXCR4 mRNA and protein in transferation group was lower than both blank group and negative control group wiht a statistical difference （P 〈 0.01 ） but there was no statistical difference between blank group and negative control group （ P 〉 0.05 ） ; （ 3 ） The cell proliferation was more rapidly in blank group and negative control group and with no statistical difference （P 〉 0.05）. The cell proliferation was de- pressed in transferation group and there was a statistical difference between the blank group or negative control group （P 〈 0.05 ） ; （4） The inhibition to invasiveness of PC-3 cells was higher in transferation group （P 〉 0.05 ） with 57.01% inhibition rate, and there was no obvious difference between the blank group or negative control group （P 〉 0.05）. CONCLUSION The expression of mRNA and protein of CXCR4 in PC-3 cells of prostate carcinoma can be inhibited by CXCR4-shRNA recombi- nant adenovirus vector and the ability of proliferation and invasiveness of cancer cell can be inhibited to some degree. It is theoretical basis and prophase conditions for animal experiment.

关 键 词：CXCR4 前列腺癌 CXCL12 RNAi 腺病毒载体 

分 类 号：Q344[生物学—遗传学]