肌动蛋白γ2对膀胱癌细胞迁移的抑制作用以及甲基化对其表达的调控作用
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摘要
目的:探讨肌动蛋白γ2(ACTG2)在膀胱癌中的表达情况,研究ACTG2对膀胱癌细胞增殖和迁移的影响,初步探讨甲基化对其的表达调控。方法:RT-PCR检测ACTG2在膀胱癌组织以及细胞系中的表达情况,构建ACTG2过表达质粒。通过MTT实验、平板克隆实验和Transwell迁移实验分别检测过表达ACTG2对膀胱癌细胞增殖、克隆形成能力以及迁移的影响。MethHc数据库预测ACTG2在膀胱癌组织中的甲基化情况,使用去甲基化药物地西他滨处理膀胱癌细胞研究甲基化对ACTG2的调控作用。结果:ACTG2在膀胱癌组织以及膀胱癌细胞系中表达水平都显著下调,过表达ACTG2可以抑制膀胱癌细胞的迁移,对膀胱癌增殖能力没有明显影响。DNA甲基化是ACTG2在膀胱癌中表达水平下降的重要原因。结论:ACTG2在膀胱癌中有抑癌功能,其表达水平受DNA甲基化调控。
Objective: To investigate the expression of actin gamma 2(ACTG2) in bladder cancer, and the effects of ACTG2 on the proliferation and migration of bladder cancer cells, and to explore the regulation of methylation on its expression.Methods: The expression of ACTG2 in bladder cancer tissues and cell lines were detected by RT-PCR, and ACTG2 overexpression plasmid was constructed.MTT assay, plate cloning assay and transwell migration assay were used to detect the effects of ACTG2 overexpression on proliferation, clone formation and migration of bladder cancer cells. MethHc database was used to predict the methylation of ACTG2 in bladder cancer tissue, and bladder cancer cells were treated with demethylated drug deoxycytidine to study the regulation effect of methylation on ACTG2.Results: ACTG2 expression was significantly down-regulated in bladder cancer tissues and bladder cancer cell lines. Overexpression of ACTG2 could inhibit the migration of bladder cancer cells and had no significant effect on the proliferation of bladder cancer. DNA methylation is an important reason for the decrease of ACTG2 expression in bladder cancer.Conclusion: ACTG2 acts as a tumor suppressor in bladder cancer, and its expression level is regulated by DNA methylation.
Objective: To investigate the expression of actin gamma 2(ACTG2) in bladder cancer, and the effects of ACTG2 on the proliferation and migration of bladder cancer cells, and to explore the regulation of methylation on its expression.Methods: The expression of ACTG2 in bladder cancer tissues and cell lines were detected by RT-PCR, and ACTG2 overexpression plasmid was constructed.MTT assay, plate cloning assay and transwell migration assay were used to detect the effects of ACTG2 overexpression on proliferation, clone formation and migration of bladder cancer cells. MethHc database was used to predict the methylation of ACTG2 in bladder cancer tissue, and bladder cancer cells were treated with demethylated drug deoxycytidine to study the regulation effect of methylation on ACTG2.Results: ACTG2 expression was significantly down-regulated in bladder cancer tissues and bladder cancer cell lines. Overexpression of ACTG2 could inhibit the migration of bladder cancer cells and had no significant effect on the proliferation of bladder cancer. DNA methylation is an important reason for the decrease of ACTG2 expression in bladder cancer.Conclusion: ACTG2 acts as a tumor suppressor in bladder cancer, and its expression level is regulated by DNA methylation.
引文
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[2] Miller KD, Siegel RL, Lin CC, et al. Cancer treatment and survivorship statistics, 2016[J]. CA Cancer J Clin, 2016,66(4):271-289.
[3] Garg M. Urothelial cancer stem cells and epithelial plasticity:current concepts and therapeutic implications in bladder cancer[J]. Cancer Metastasis Rev, 2015,34(4):691-701.
[4] Lamy A, Gobet F, Laurent M, et al. Molecular profiling of bladder tumors based on the detection of FGFR3and TP53 mutations[J]. J Urol, 2006, 176(6 Pt 1):2 686-2 689.
[5] Urakami S, Shiina H, Enokida H, et al. Epigenetic inactivation of Wnt inhibitory factor-1 plays an important role in bladder cancer through aberrant canonical Wnt/beta-catenin signaling pathway[J]. Clin Cancer Res,2006,12(2):383-391.
[6] Thorson W, Diaz-horta O, Foster J 2nd, et al. De novo ACTG2 mutations cause congenital distended bladder, microcolon, and intestinal hypoperistalsis[J].Hum Genet, 2014,133(6):737-742.
[7] Drew JE, Farquharson AJ, Mayer CD, et al. Predictive gene signatures:molecular markers distinguishing colon adenomatous polyp and carcinoma[J]. PLoS One, 2014,9(11):e113071.
[8] Demicco EG, Boland GM, Brewer Savannah KJ, et al. Progressive loss of myogenic differentiation in leiomyosarcoma has prognostic value[J]. Histopathology,2015,66(5):627-638.
[9] Edfeldt K, Hellman P, Westin G, et al. A plausible role for actin gamma smooth muscle 2(ACTG2)in small intestinal neuroendocrine tumorigenesis[J]. BMC Endocr Disord, 2016,16:19.
[10] Zhang C, Zhao H, Li J, et al. The identification of specific methylation patterns across different cancers[J].PLoS One, 2015,10(3):e0120361.
[11] Chen W, Zheng R, Baade PD, et al. Cancer statistics in China, 2015[J]. CA Cancer J Clin, 2016,66(2):115-132.
[12] Watson MB, Lind MJ, Smith L, et al. Expression microarray analysis reveals genes associated with in vitro resistance to cisplatin in a cell line model[J]. Acta Oncol, 2007,46(5):651-658.
[13] Schulz WA, Goering W. DNA methylation in urothelial carcinoma[J]. Epigenomics, 2016, 8(10):1 415-1 428.
[14] Rani A, Greenlaw R, Smith RA, et al. HES1 in immunity and cancer[J]. Cytokine Growth Factor Rev,2016,30:113-117.
[15] Huang XY, Gan RH, Xie J, et al. The oncogenic effects of HES1 on salivary adenoid cystic carcinoma cell growth and metastasis[J]. BMC Cancer, 2018, 18(1):436.