下调SPOCK1基因表达可抑制膀胱癌细胞的增殖、迁移和侵袭

Down-regulation of SPOCK1 gene expression inhibits the proliferation, migration and invasion of bladder cancer cells

目的:检测膀胱癌细胞中SPOCK1(sparc/osteonectin,cwcv and kazal-like domains proteoglycan 1)基因的表达情况,并进一步研究下调SPOCK1基因表达对膀胱癌细胞增殖、迁移和侵袭的影响。方法 :采用实时荧光定量PCR和蛋白质印迹法检测人输尿管上皮永生化细胞SV-HUC-1以及膀胱癌细胞株5637和T24中SPOCK1基因的表达情况。构建特异性针对SPOCK1基因的si RNA片段,并将其通过脂质体介导转染至膀胱癌5637细胞;同时设立未转染的空白对照组和转染无义序列的阴性对照组。采用实时荧光定量PCR法和蛋白质印迹法检测各组细胞中SPOCK1 m RNA和蛋白的表达差异。然后采用CCK-8法、划痕愈合实验和Transwell小室法分别检测SPOCK1基因表达下调后膀胱癌细胞增殖、迁移和侵袭能力的变化情况。结果 :膀胱癌细胞中SPOCK1基因表达水平高于输尿管上皮永生化细胞,尤以膀胱癌5637细胞中的表达水平最高(P值均<0.05)。SPOCK1 si RNA转染后,膀胱癌5637细胞中SPOCK1基因的表达明显下调(P<0.05),而膀胱癌细胞的增殖、迁移和侵袭能力均被明显抑制(P值均<0.05)。结论:膀胱癌细胞中SPOCK1基因存在过表达现象。下调SPOCK1基因表达可以降低膀胱癌5637细胞的增殖、迁移和侵袭能力。

Objective： To detect the expression of sparc/osteonectin, cwcv and kazal-like domains proteoglycan 1（SPOCK1） gene in bladder cancer cells, and further to investigate the effects of down-regulation of SPOCK1 gene expression on the proliferation, migration and invasion of bladder cancer cells.Methods： The expression level of SPOCK1 gene in human ureteralepithelial immortalized cell line SV-HUC-1 and bladder cancer cell lines 5637 and T24 was detected by real-time fluorescent quantitative PCR（RFQ-PCR） and Western blotting, respectively. The si RNA fragments targeting SPCOK 1 gene were constructed and transfected into bladder cancer 5637 cells by liposome（as the experimental group）; at the same time, the 5637 cells were transfected without anything as the blank control group and transfected with scramble si RNA as the negative control group. Then the expressions of SPOCK1 m RNA and protein in each group were detected by RFQ-PCR and Western blotting, respectively. The changes of proliferation, migration and invasion abilities of bladder cancer 5637 cells after SPOCK1 expression down-regulation were detected by CCK-8 method, Wound healing assay and Transwell chamber assay, respectively. Results： As compared with the ureteral epithelial immortalized cells, the expression levels of SPOCK1 m RNA and protein were higher in bladder cancer cells, especially in 5637 cells（both P〈 0.05）. After transfection with SPOCK1 si RNA, the expression of SPOCK1 gene was downregulated（P 〈0.05）, the proliferation, migration and invasion of bladder cancer 5637 cells were significantly inhibited（all P 〈0.05）.Conclusion： SPOCK1 gene is overexpressed in bladder cancer cells. The down-regulation of SPOCK1 gene expression can reduce the proliferation, migration and invasion of bladder cancer 5637 cells.