下调脂肪酸合成酶表达对膀胱癌UMUC3细胞增殖、迁移及侵袭能力的影响

Effect of down-regulation of fatty acid synthase expression on proliferation,migration and invasion of bladder carcinoma UMUC3 cell lines

目的:研究脂肪酸合成酶(FASN)表达对膀胱癌UMUC3细胞增殖、迁移、侵袭的影响,探讨其内在可能机制。方法:免疫组化法检测30例膀胱癌和15例正常膀胱组织FASN蛋白的表达;用脂质体2000分别转染FASN siRNA和无义siRNA至UMUC3细胞,筛选、鉴定siFASN和siControl稳定的细胞,siFASN组细胞设为实验组,siControl组设为对照组;采用蛋白印迹法(Western blot)和实时荧光定量PCR(RT-PCR)法分别检测siFASN组和siControl组细胞FASN蛋白及mRNA的表达,MTT法检测siFASN组和siControl组细胞增殖情况,划痕试验、Transwell试验分别检测siFASN组和siControl组细胞迁移、侵袭能力。结果:FASN蛋白在膀胱癌组织中过表达,且与病理分期、分级密切相关(P<0.05)。与siControl组相比,siFASN组细胞FASN mRNA及蛋白表达下调(P<0.05),细胞增殖活力明显下降(P<0.05),迁移能力明显下降(P<0.05),穿膜细胞数量明显减少(P<0.05)。结论:FASN过表达在膀胱癌发生、发展中发挥重要作用,下调FASN表达能抑制膀胱癌细胞的增殖、迁移、侵袭能力,抑制FASN表达有望成为一种新的膀胱癌治疗方法。

Objective:To investigate the effects of fatty acid synthase(FASN)on proliferation,migration and invasion of bladder cancer UMUC3 cell lines and possible mechanism.Methods:The expression levels of FASN protein in 30 cases of bladder cancer and 15 cases of normal bladder tissues were detected by Immunohistochemistry.FASN siRNA and nonsense siRNA were transfected into UMUC3 cell lines by lipofectamine 2000 respectively,and the stable siFASN and siControl cell lines were successfully obtained after screening and identification for several times.The siFASN cell lines were set as the experimental group,while the siControl cell lines were set as the control group.The expressions of FASN protein and mRNA in the experimental group and the control group were detected by Western blot and real-time quantitative PCR(RT-PCR)respectively.Cell proliferation activities in two groups were detected by MTT assay and cell invasion and migration in two groups were detected by cell scratch test and Transwell invasive assays respectively.Results:FASN protein was overexpressed in bladder cancer tissues,and it was closely correlated with pathological stage and grade(P<0.05).Compared with the siControl group,the expressions of FASN mRNA and protein in the siFASN group cell lines were decreased significantly(P<0.05).The cell proliferation ability,the migration ability and the number of transmembrane cells of siFASN group cell lines were reduced significantly(P<0.05).Conclusion:The FASN overexpression may play an essential role in the development and progression of bladder cancer.Down-regulation of FASN expression can inhibit the proliferation,migration and invasion of bladder cancer cells,and inhibition of FASN expression is expected to be a new treatment for bladder cancer.