肺癌抑癌基因1对人前列腺癌T_3B细胞增殖的抑制效应

The proliferation inhibition effects of TSLC1 gene on human prostatic carcinoma cell line T_3B

背景与目的:肺癌抑癌基因1(tumor suppressor in lung cancer1,TSLC1)是新近发现的一种肿瘤抑制基因,它在包括前列腺癌在内的多种恶性肿瘤中是失活的。本研究通过将TSLC1转染至人前列腺癌T3B细胞来观察T3B细胞增殖的受抑情况。方法:将克隆有TSLC1全长cDNA的真核重组载体pCI-TSLC1稳定转染至前列腺癌T3B细胞中(实验组)。以转染空质粒pCI-neo的T3B细胞为对照组,野生型T3B细胞为空白组。四甲基偶氮唑蓝法检测细胞增殖,FACSort流式细胞仪检测细胞周期,AnnexinV/PI双染法检测细胞凋亡情况。结果:实验建立了高表达TSLC1蛋白的稳定细胞株。与对照组和空白组相比,实验组细胞生长速度减慢,增殖受到明显抑制,G0/G1期细胞为(80±3)%,高于对照组(66±4)%和空白组(64.2±0.9)%;S期细胞为(11.1±1.3)%,低于对照组(24.0±2.4)%和空白组(26.2±0.6)%,组间差异有显著性(P<0.01),实验组细胞周期发生了明显的G0/G1期阻滞。实验组细胞早期凋亡率,晚期凋亡率和总凋亡率分别为(11.9±0.4)%,(10.2±0.4)%和(22.1±0.6)%,与对照组和空白组细胞相比均明显升高(P<0.01)。结论:TSLC1基因明显抑制T3B细胞增殖,并诱导细胞发生凋亡。

Background and purpose： The TSLCl（tumor suppressor in lung cancer 1） gene has recently been identified as a novel tumor suppressor, and it is inactivated in various cancers including prostate carcinoma, etc. In this study, we transfected TSLC1 into human prostate carcinoma cell line T3B to observe the proferative inhibition of T3B cells. Methods： Enkaryotic vector pCI-TSLC 1 that had full length of TSLC 1 cDNA were transfected into human prostate carcinoma cell line T3B. The T3B cells transfected with the plasmid （experimental group） and those treated with pCI-neo vector（control group） and without any treatment （blank group） were compared. Cell proliferation was analyzed with MTT assay. FACSort flow cytometry analysis was performed to assess the cell cycle distribution and apoptosis. Results： A stable cell line expressing TSLC 1 protein was successfully established. The growth of TSLC 1-transfected cells was significantly suppressed in vitro compared with those of the control and blank groups. The amount of G0/GI cells was （80±3）% in the experimental group, while those of the control and blank groups were （66±4）% and （64.2±0.9）% respectively. The amount of S phase cells in experimental group, 11.1%±1.3%, was significantly lower（P〈0.01） than that of the control group（24.0%±2.4%） and the blank group（26.2%±0.6%）, which suggested a G0/G1 cell cycle arrest. The number of cells in early, late and total phase apoptosis （11.9%±0.4%, 10.2%±0.4% and 22.1%±0.6% respectively） were significantly higher than those of the control and blank groups（P〈0.01）. Conclusion： TSLC1 strongly inhibits the proliferation of T3B cells in vitro and induces apoptosis of prostate carcinoma cells.