Yin Shanlan, Zhang Quanle, Wang Yuhong, Li Shaoru, Hu Ruili
Department of Gynecology and Obstetrics, The First Affiliated Hospital of Xinxiang Medical University, Weihui, Henan 453100, P.R. China.
Exp Ther Med. 2019 Apr;17(4):2837-2846. doi: 10.3892/etm.2019.7226. Epub 2019 Jan 31.
Human papillomaviruses (HPVs) have important roles in the development and progression of cervical cancer, but the underlying mechanisms are yet to be fully elucidated. MicroRNA-130a (miR-130a) has previously been reported to promote cervical cancer growth. However, the underlying molecular mechanisms by which miR-130a promotes cervical cancer progression have remained largely elusive. In the present study, polymerase chain reaction and western blot analyses were performed to examine the expression levels of miR-130a and associated proteins. A wound healing assay and a Transwell assay were applied to study cell migration and invasion. A luciferase reporter gene assay was performed to confirm the targeting associations of miR-130a. It was observed that miR-130a was significantly upregulated in cervical cancer tissues compared with that in adjacent non-tumorous tissues. High expression of miR-130a was significantly associated with lymph node metastasis and an advanced clinical stage of cervical cancer. Furthermore, the expression of miR-130a was also higher in HPV(+) cervical cancer cell lines compared with that in HPV(-) cells. Knockdown of HPV18 E6 significantly inhibited the expression of miR-130a in HeLa cervical cancer cells. Furthermore, knockdown of miR-130a reduced the migration and invasion of HeLa cells. Tissue inhibitor of metalloproteinase 2 (TIMP2), an antagonist of matrix metalloproteinase 2 (MMP2), was identified as a novel, direct target gene of miR-130a. The expression of TIMP2 was negatively mediated by miR-130a, and HPV18 E6 inhibited the expression of TIMP2 in HeLa cells. Furthermore, knockdown of TIMP2 rescued the suppressive effects of miR-130a downregulation on the migration and invasion of HeLa cells. In summary, the present study suggests that HPV18 E6 promotes the expression of miR-130a, which further inhibits the expression of TIMP2 and promotes cervical cancer cell invasion. Therefore, HPV/miR-130a/TIMP2 signaling may be a potential target for the prevention of cervical cancer metastasis.
人乳头瘤病毒（HPV）在宫颈癌的发生和发展中起着重要作用，但其潜在机制尚未完全阐明。先前有报道称，微小RNA - 130a（miR - 130a）可促进宫颈癌生长。然而，miR - 130a促进宫颈癌进展的潜在分子机制在很大程度上仍不清楚。在本研究中，进行了聚合酶链反应和蛋白质印迹分析，以检测miR - 130a及相关蛋白的表达水平。采用伤口愈合试验和Transwell试验研究细胞迁移和侵袭。进行荧光素酶报告基因试验以确认miR - 130a的靶向关联。结果发现，与相邻非肿瘤组织相比，宫颈癌组织中miR - 130a显著上调。miR - 130a的高表达与宫颈癌的淋巴结转移和临床晚期显著相关。此外，与HPV（-）细胞相比，HPV（+）宫颈癌细胞系中miR - 130a的表达也更高。敲低HPV18 E6可显著抑制HeLa宫颈癌细胞中miR - 130a的表达。此外，敲低miR - 130a可减少HeLa细胞的迁移和侵袭。金属蛋白酶组织抑制剂2（TIMP2）是基质金属蛋白酶2（MMP2）的拮抗剂，被确定为miR - 130a的一个新的直接靶基因。TIMP2的表达受到miR - 130a的负向调节，HPV18 E6抑制HeLa细胞中TIMP2的表达。此外，敲低TIMP2可挽救miR - 130a下调对HeLa细胞迁移和侵袭的抑制作用。总之，本研究表明HPV18 E6促进miR - 130a的表达，进而抑制TIMP2的表达并促进宫颈癌细胞侵袭。因此，HPV/miR - 130a/TIMP2信号通路可能是预防宫颈癌转移的一个潜在靶点。
