摘要
目的探讨SLP-2(stomatin-like protein 2)基因对人前列腺癌PC-3细胞增殖和凋亡的影响。方法合成和构建SLP-2沉默和过表达载体,用LipofectamineTM 2000分别转染5组人前列腺癌PC-3细胞:pcDNA3.1-SLP-2组,空质粒pcDNA3.1组,siRNA-SLP-2组,siRNA阴性对照组以及空白对照组。采用Q-PCR和Western-Blot分别检测PC-3细胞中SLP-2的mRNA和蛋白表达量;用细胞计数试剂盒(CCK-8)检测PC-3细胞增殖;采用细胞线粒体膜电位JC-1试剂盒检测PC-3细胞的早期凋亡情况。结果转染pcDNA3.1-SLP-2质粒和siRNA-SLP-2后PC-3细胞SLP-2基因mRNA表达水平分别为(324.884±16.508)和(0.195±0.016),蛋白表达水平分别为(5.013±0.284)和(0.296±0.011),与空白对照组相比差异均有统计学意义(P<0.05);转染pcDNA3.1-SLP-2基因48 h、72 h、96 h后PC-3细胞增殖吸光度值(OD值)分别为(0.714±0.007)、(1.103±0.018)、(1.348±0.018),均显著高于空白对照组(P<0.05)。采用siRNA沉默SLP-2基因48 h、72 h、96 h后PC-3细胞增殖率分别为(0.571±0.004)、(0.875±0.010)、(1.044±0.008),均显著低于空白对照组(P<0.05);细胞凋亡结果显示,转染pcDNA3.1-SLP-2和siRNA-SLP-2的PC-3细胞早期凋亡率分别为(2.433±0.577)和(9.197±0.602),与空白对照组(4.993±0.710)相比差异均有统计学意义(P<0.05)。结论 SLP-2可促进人前列腺癌PC-3细胞的增殖,并抑制细胞的早期凋亡。
Objective To explore the effect of SLP-2 on cell proliferation and early apoptosis in PC-3 cells line of prostate cancer. Methods The pcDNA3.1-SLP-2 and siRNA-SLP-2 were constructed and transfected into PC-3 cells by LipofectamineTM 2000 in 5 groups： pcDNA3.1-SLP-2, pcDNA3.1, siRNA-SLP-2, mock siRNA, and blank control. Q-PCR and Western blot were used to detect the expression of SLP-2 mRNA and protein, CCK-8 kit was used to detect cell proliferation, and MMP （mitochondrial membrane potential） JC-1 kit for cell apoptosis determination. Results The mRNA expression levels of SLP-2 in pcDNA3.1-SLP-2 and siRNA-SLP-2 were （324.884±16.508） and （0.195±0.016）, respectively. The protein expression levels were （5.013±0.284） and （0.296±_0.011）, respectively. There were statistical significance in both mRNA and protein expression levels of SLP-2 between these two groups and blank control group （P〈0.05）. The OD values in pcDNA3.1-SLP-2 group were （0.714±0.007）, （1.103±0.018）, （1.348±0.018）, respectively, at 48, 72, 96 h after transfection, which were significantly higher than those of the blank control. The OD values in siRNA-SLP-2 group were （0.571±0.004）, （0.875±0.010）, （1.044±0.008）,respectively, at 48, 72, 96 h after transfection, which were significantly lower than those of the blank control. The cell apoptosis assay showed that the early apoptosis rates of pcDNA3.1-SLP-2 and siRNA-SLP-2 were （2.433±0.577） and （9.197±0.602）, respectively. There were statistical significance in the early apoptosis rates between these two groups and blank control group （P〈0.05）. Conclusion SLP-2 could promote cell proliferation and inhibit the early cell apoptosis of PC-3 cell line.
出处
《中华腔镜泌尿外科杂志（电子版）》
2014年第1期50-53,共4页
Chinese Journal of Endourology(Electronic Edition)
基金
国家自然科学基金(81001139
81202012)
关键词
SLP-2
前列腺癌
PC-3细胞
增殖
凋亡
Stomatin-like protein 2
Prostate cancer, PC-3 cells
Proliferation
Apoptosis
