miR-206通过调控PDCD10抑制前列腺癌细胞增殖、侵袭和迁移

The miR-206 inhibits proliferation，migration and invasion of prostate cancer cells by targeting PDCD10

摘要:

目的：探讨miR-206通过调控程序性细胞死亡因子10（programmed cell death 10，PDCD10）对前列腺细胞增殖、侵袭和迁移的影响。方法：选取国药汉江医院手术切除的前列腺癌标本61例，另选40例良性前列腺增生组织作为对照，采用原位杂交检测miR-206和PDCD10表达水平，分别分析miR-206和PDCD10的表达与前列腺癌患者临床病理参数间的关系；双荧光素酶实验检测miR-206和PDCD10在前列腺癌细胞中的相互作用；选取前列腺癌PC3细胞，分别转染miR-206 mimics+PDCD10 pcDNA过表达载体、miR-206 mimics、PDCD10 pcDNA过表达载体、mimics NC组和mimics NC+PDCD10 pcDNA组。采用实时荧光定量PCR法检测PDCD10 mRNA的表达量，Western blot检测PDCD10蛋白的表达量；通过MTT、Transwell迁移和侵袭检测前列腺癌细胞的增殖、迁移和侵袭能力。结果：原位杂交实验染色显示，miR-206在前列腺癌组织中的阳性表达率（24.59%，15/61）明显低于良性前列腺增生组织的阳性表达率（87.5%，35/40）（ χ2=38.248，P=0.000）。PDCD10在前列腺癌组织中的阳性表达率（73.77%，45/61）明显高于良性前列腺增生组织的阳性表达率（27.5%，11/40）（ χ2=20.397，P=0.000）。miR-206与PDCD10均与TNM临床分期、Gleason评分等有关（P<0.05），与患者年龄、术前PSA水平等均无关（P >0.05）。线性回归与相关性分析发现，miR-206表达与PDCD10表达呈明显的负相关性（r=-0.9594，P<0.001）。miR-206 mimic与PDCD10野生型报告载体共转后，细胞中荧光素酶的活性降低（P<0.01）。miR-206 mimics组PDCD10 mRNA和蛋白表达水平最低。过表达miR-206可显著抑制PC3细胞的增殖、迁移和侵袭能力。过表达PDCD10可显著提高PC3细胞的增殖、迁移和侵袭能力，且过表达PDCD10可逆转miR-206对PC3细胞增殖、迁移和侵袭能力的抑制作用。结论：miR-206可抑制前列腺癌细胞增殖、迁移和侵袭，其作用机制与靶向负调控PDCD10的表达有关。

Abstract:

Objective：To investigate the effect of miR-206 on proliferation，invasion and migration of prostate cells by regulating pro－grammed cell death 10（PDCD10）. Methods：Prostate cancer specimens of 61 cases in Sinopharm Hanjiang Hospital were selected and 40 benign prostatic hyperplasia tissues were selected as controls. Expression levels of miR-206 and PDCD10 were detected by in situ hybridization，and the relationship between expressions of miR-206 or PDCD10 and clinicopathological parameters of patients with prostate cancer was analyzed. The interaction between miR-206 a and PDCD10 were verified by dual luciferase reporter gene assay. Prostate cancer PC3 cells were transfected into miR-206 mimics+PDCD10 pcDNA overexpression vector，miR-206 mimics，PDCD10 pcDNA overexpression vector，the mimics NC group and the mimics NC+PDCD10 pcDNA group，respectively. The expres－sion of PDCD10 mRNA was detected by RT-qPCR and the expression of PDCD10 protein was detected by Western blot. The proliferation，migration and invasion of prostate cancer cells were detected by MTT and Transwell. Results：In situ hy－bridization showed that the positive expression rate of miR-206 in prostate cancer tissues（24.59%，15/61） was significantly lower than that in benign prostatic hyperplasia tissues（87.5%，35/40）（χ2=38.248，P=0.000）. The positive expression rate of PDCD10 in prostate cancer tissues（73.77%，45/61） was significantly higher than that in benign prostatic hyperplasia tissues（27.5%，11/40）（χ2=20.397，P=0.000）. Both miR-206 and PDCD10 were associated with TNM clinical stage and Gleason score（P<0.05），and were not associated with age and preoperative PSA levels（P>0.05）. Linear regression and correlation analysis showed that miR-206 expression had a significant negative correlation with PDCD10 expression（r=-0.959，P<0.001）. After co-transfection of miR-206 mimic with the PDCD10 wild-type reporter vector，the luciferase activity in cells was decreased（P<0.01）. The expression level of PDCD10 mRNA and protein was the lowest in the miR-206 mimics group. Overexpression of miR-206 significantly inhibited the proliferation，migration and invasion of PC3 cells. Overexpression of PDCD10 was able to significantly increase the proliferation，migration and invasion of PC3 cells，and overexpression of PDCD10 was able to reverse the inhibitory effect of miR-206 on the cell proliferation，migration and invasion abilities of PC3 cells. Conclusion：The miR-206 can inhibit the proliferation，migration and invasion of prostate cancer cells，whose mechanism is related with the targeted negative regulation of PDCD10 expression.