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作 者:罗黔[1] 吕杰[1] 王宁[1] LUO Qian;LV Jie;WANG Ning(Department of Urology,Luzhou People's Hospital,Luzhou 646000,Sichuan Province,China)
机构地区:[1]泸州市人民医院泌尿外科,四川泸州646000
出 处:《中国临床解剖学杂志》2020年第3期295-300,共6页Chinese Journal of Clinical Anatomy
基 金:四川省科技厅重点项目(2015JY0224)。
摘 要:目的探讨MALAT1-mir-30a-5p-SOX4轴在膀胱癌中的调控机制,以期阐明膀胱癌的分子机制和潜在的治疗靶点。方法RT-PCR检测mir-30a-5p、MALAT1及SOX4在膀胱癌细胞中的表达;荧光素酶报告检测验证mir-30a-5p和SOX4之间的生物学关系。通过体外实验研究mir-30a-5p和SOX4在T24细胞中的生物学功能,包括CCK-8法检测细胞增殖情况,Transwell检测细胞侵袭,划痕法检测细胞迁移;Western blot检测蛋白表达。结果在膀胱癌细胞中,MALAT1的表达上调,mir-30a-5p表达下调(P<0.01);而sh-MALAT1抑制T24细胞增殖、侵袭和迁移(P<0.01)。进一步的研究表明,MALAT1可以直接与mir-30a-5p相互作用,且SOX4是mir-30a-5p的靶点,SOX4可以被mir-30a-5p过表达下调(P<0.01)。结论敲除MALAT1可解除MALAT1对mir-30a-5p的抑制从而抑制SOX4,MALAT1可能通过调节miR-30a/SOX4通路调节膀胱癌的增殖、侵袭和转移。Objective To elucidate the molecular mechanism and potential therapeutic targets of bladder cancer and explore the regulatory mechanism of MALAT1-miR-30 a-5 p-SOX4 axis in bladder cancer.Methods Quantitative real-time PCR(RT-PCR)was used to detect the expressions of mir-30 a-5 p,MALAT1 and SOX4 in bladder cancer cells.Luciferase reporter assay was used to verify the biological relationship between mir-30 a-5 p and SOX4.The biological functions of mir-30 a-5 p and SOX4 in T24 cells were studied by in vitro experiments,including the detection of cell proliferation by CCK-8 method,cell invasion by Transwell and cell migration by wound healing method.Western blot was used to detect protein expression.Results In bladder cancer cells,MALAT1 expression was up-regulated and mir-30 a-5 p expression was down-regulated(P<0.01).Sh-MALAT1 inhibited the proliferation,invasion and migration of T24 cells.Further studies showed that MALAT1 can directly interact with mir-30 a-5 p,and SOX4 is the target of miR-30 a-5 p,and SOX4 can be down-regulated by over-expression mir-30 a-5 p.Conclusions MALAT1 knockout can relieve the inhibition of mir-30 a-5 p by MALAT1 and thus inhibiting SOX4.MALAT1 may regulate the proliferation,invasion and metastasis of bladder cancer by regulating the mir-30 a/SOX4 pathway.
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