Liu Zhi-Jie, Liu Shi-Hui, Li Jun-Ru, Bie Xiao-Chuan, Zhou Yang
Department of Urology, Hanting District People's Hospital of Weifang, Weifang, Shandong 261100, People's Republic of China.
Cancer Manag Res. 2020 Oct 28;12:10669-10678. doi: 10.2147/CMAR.S266421. eCollection 2020.
In order to investigate the role of miR-15b-5b in the progression of prostate cancer.
We employed RT-qPCR assay to analyze the transcriptional level of miR-15b-5b in cell lines including PC-3, prostate cancer tissues as well as normal prostate tissues. The protein level of large tumor suppressor factor 2 (LATS2) was detected by Western blot in similar specimens. Bioinformatic analysis was used to predict the targets of miR-15b-5p, and dual-luciferase assay was performed to confirm the relationship of miR-15b-5p with LATS2. Cell proliferation assay and colony formation assay were used to assess the effects of miR-15b-5b on the proliferation of PC-3 cells. Multivariate analysis was performed to identify factors associated with overall survival using the Cox proportional hazards model.
MiR-15b-5b was up-regulated in prostate cancer tissues as well as cell lines, and increased expression of miR-15b-5b was highly correlated with the poor prognosis of patients with prostate cancer. Ectopic expression of miR-15b-5b promoted the proliferation of PC-3 cells. Reciprocally, silence of miR-15b-5b elicited opposite effects on cell proliferation. Mechanistically, we identified LATS2 as the target of miR-15b-5b, which in turn limited LATS2 expression in PC-3 cells. Furthermore, the stimulatory effects of miR-15b-5b on cell proliferation can be attenuated by overexpression of LATS2. Conversely, inhibition of LATS2 promoted the proliferation of PC-3 cells induced by miR-15b-5b. Our data thus demonstrate that dysregulation of miR-15b-5b exacerbates prostate cancer progression via suppression of LATS2.
The identification of the oncogenic role of miR-15b-5b in prostate cancer thus proposes that miR-15b-5p might be a new therapeutic target for the treatment of prostate cancer.
探讨miR-15b-5b在前列腺癌进展中的作用。
我们采用RT-qPCR检测法分析miR-15b-5b在包括PC-3细胞系、前列腺癌组织以及正常前列腺组织中的转录水平。通过蛋白质免疫印迹法检测类似标本中肿瘤抑制因子2(LATS2)的蛋白水平。利用生物信息学分析预测miR-15b-5p的靶标,并进行双荧光素酶测定以确认miR-15b-5p与LATS2的关系。采用细胞增殖检测和集落形成检测评估miR-15b-5b对PC-3细胞增殖的影响。使用Cox比例风险模型进行多因素分析,以确定与总生存相关的因素。
miR-15b-5b在前列腺癌组织和细胞系中上调,且miR-15b-5b表达增加与前列腺癌患者的不良预后高度相关。miR-15b-5b的异位表达促进了PC-3细胞的增殖。相反,miR-15b-5b沉默对细胞增殖产生相反的影响。机制上,我们确定LATS2是miR-15b-5b的靶标,这反过来限制了PC-3细胞中LATS2的表达。此外,LATS2的过表达可减弱miR-15b-5b对细胞增殖的刺激作用。相反,抑制LATS2可促进miR-15b-5b诱导的PC-3细胞增殖。因此,我们的数据表明,miR-15b-5b的失调通过抑制LATS2加剧前列腺癌进展。
miR-15b-5b在前列腺癌中的致癌作用的鉴定表明,miR-15b-5p可能是治疗前列腺癌的新治疗靶点。