Huang Fu, Zhou Li Quan
Department of Urology, The Second Affiliated Hospital of Guangxi Medical University, China.
Appl Bionics Biomech. 2022 Jun 6;2022:9292538. doi: 10.1155/2022/9292538. eCollection 2022.
To investigate the effect of long noncoding RNA (lncRNA) CERS6 antisense RNA1 (CERS6-AS1) on the biological behavior of prostate cancer cells DU145 and its mechanism.
RT-PCR was used to detect the relative level of CERS6-AS1 and miR-16-5p in prostate cancer tissues, adjacent tissues, prostate cancer cells DU145, and human normal prostate epithelial cells RWPE-1. DU145 cells were divided into control group, si-CERS6-AS1 group, si-NC group, miR-16-5p mimic group, miR-NC group, and si-CERS6-AS1+miR-16-5p inhibitor group. And CCK-8 method and colony formation test was applied to detect cell proliferation ability, flow cytometry was selected to calculate cell apoptosis, and scratch healing test was employed to assess cell migration ability. Western blot was determined to detect high mobility protein A2 (HMGA2) expression. RT-PCR and dual-luciferase reporter experiments were used to analyze the targeting relationship among CERS6-AS1, miR-16-5p, and HMGA2.
Compared with the adjacent tissues, the relative level of CERS6-AS1 in prostate cancer tissue was increased ( < 0.05), and the relative level of miR-16-5p was decreased ( < 0.05). Compared with RWPE-1 cells, the relative level of CERS6-AS1 in DU145 cells was increased ( < 0.05), and the relative level of miR-16-5p was decreased ( < 0.05). Compared with the control group and the si-NC group, the HMGA2 protein expression, the colony formation number, and the scratch healing rate of DU145 cells in the si-CERS6-AS1 group and the miR-16-5p mimic group were reduced ( < 0.05), and the relative level of miR-16-5p and the proliferation inhibition rate and apoptosis were increased ( < 0.05). miR-16-5p is specifically bound to CERS6-AS1 and HMGA2, respectively. Compared with the si-CERS6-AS1 group, the HMGA2 protein expression, the colony formation number, and the scratch healing rate of DU145 cells in the si-CERS6-AS1+miR-16-5p inhibitor group were increased ( < 0.05), and the cell proliferation inhibition rate and apoptosis rate were reduced ( < 0.05).
Silencing CERS6-AS1 can inhibit the proliferation and migration of prostate cancer cell DU145 and induce cell apoptosis, the mechanism is related to the regulation of the miR-16-5p/HMGA2 axis.
探讨长链非编码RNA（lncRNA）CERS6反义RNA1（CERS6-AS1）对前列腺癌细胞DU145生物学行为的影响及其机制。
采用RT-PCR检测前列腺癌组织、癌旁组织、前列腺癌细胞DU145及人正常前列腺上皮细胞RWPE-1中CERS6-AS1和miR-16-5p的相对水平。将DU145细胞分为对照组、si-CERS6-AS1组、si-NC组、miR-16-5p模拟物组、miR-NC组和si-CERS6-AS1+miR-16-5p抑制剂组。应用CCK-8法和集落形成试验检测细胞增殖能力，采用流式细胞术计算细胞凋亡率，采用划痕愈合试验评估细胞迁移能力。通过蛋白质免疫印迹法检测高迁移率族蛋白A2（HMGA2）的表达。采用RT-PCR和双荧光素酶报告基因实验分析CERS6-AS1、miR-16-5p和HMGA2之间的靶向关系。
与癌旁组织相比,前列腺癌组织中CERS6-AS1的相对水平升高（<0.05），miR-16-5p的相对水平降低（<0.05）。与RWPE-1细胞相比，DU145细胞中CERS6-AS1的相对水平升高（<0.05），miR-16-5p的相对水平降低（<０.05）。与对照组和si-NC组相比，si-CERS6-AS1组和miR-16-5p模拟物组中DU145细胞的HMGA2蛋白表达、集落形成数和划痕愈合率降低（<0.05），miR-16-5p的相对水平、增殖抑制率和凋亡率升高（<0.05）。miR-16-5p分别与CERS6-AS1和HMGA２特异性结合。与si-CERS6-AS1组相比，si-CERS6-AS1+miR-16-5p抑制剂组中DU145细胞的HMGA2蛋白表达、集落形成数和划痕愈合率升高（<0.05）,细胞增殖抑制率和凋亡率降低（<0.05）。
沉默CERS6-AS1可抑制前列腺癌细胞DU145的增殖和迁移并诱导细胞凋亡，其机制与调控miR-16-5p/HMGA2轴有关。
