PRR34-AS1 通过海绵吸附 miR-498 促进 TOMM20 和 ITGA6 介导的 HCC 肿瘤进展。
PRR34-AS1 sponges miR-498 to facilitate TOMM20 and ITGA6 mediated tumor progression in HCC.
机构信息
Department of Medical Oncology, Shanxi Bethune Hospital& Shanxi Academy of Medical Sciences, Taiyuan 030032, China.
Department of Gastroenterology, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, China.
出版信息
Exp Mol Pathol. 2021 Jun;120:104620. doi: 10.1016/j.yexmp.2021.104620. Epub 2021 Feb 17.
BACKGROUND
The researches on PRR34 antisense RNA 1 (PRR34-AS1) have been limited. Both translocase of outer mitochondrial membrane 20 (TOMM20) and integrin subunit alpha 6 (ITGA6) have been proven to facilitate cancer progression. Whether TOMM20 or ITGA6 affects hepatocellular carcinoma (HCC) progression has never been investigated. Some studies showed that microRNA 498 (miR-498) can suppress HCC progression. Additionally, the influence of ceRNA network (including PRR34-AS1, miR-498, and TOMM20 or ITGA6) on HCC progression has not been inquired into yet.
METHODS
The knockdown or overexpression efficiency was validated via RT-qPCR. Also, RT-qPCR was applied to detect the expression of PRR34-AS1, miR-498, TOMM20, and ITGA6. Cell proliferation in HCC was tested via EdU and colony formation assays. Transwell assays presented the migratory and invasive capabilities of HCC cells. Subcellular fractionation and FISH assays showed the subcellular localization of PRR34-AS1. RNA pull down and luciferase reporter assays were performed to explore whether miR-498 combines with PRR34-AS1, TOMM20 or ITGA6. Western blot was conducted to detect protein expression. Rescue experiments were conducted to verify the relationship among PRR34-AS1, miR-498, TOMM20, and ITGA6.
RESULTS
The expressions of PRR34-AS1, TOMM20, and ITGA6 were markedly high in HCC cell lines while miR-498 was lowly expressed. PRR34-AS1, TOMM20, and ITGA6 promoted HCC progression while miR-498 suppressed cell proliferation, migration, and invasion in HCC. Furthermore, PRR34-AS1, TOMM20, and ITGA6 combined with miR-498.
CONCLUSION
PRR34-AS1 facilitates HCC progression by regulating miR-498/TOMM20/ITGA6 axis.
背景
PRR34 反义 RNA1（PRR34-AS1）的研究受到限制。已经证实外膜转位酶 20（TOMM20）和整合素亚基α6（ITGA6）有助于癌症进展。TOMM20 或 ITGA6 是否影响肝细胞癌（HCC）进展尚未被研究。一些研究表明 microRNA 498（miR-498）可以抑制 HCC 进展。此外，还没有研究 ceRNA 网络（包括 PRR34-AS1、miR-498、TOMM20 和 ITGA6）对 HCC 进展的影响。
方法
通过 RT-qPCR 验证敲低或过表达效率。还应用 RT-qPCR 检测 PRR34-AS1、miR-498、TOMM20 和 ITGA6 的表达。通过 EdU 和集落形成测定检测 HCC 中的细胞增殖。Transwell 测定显示 HCC 细胞的迁移和侵袭能力。亚细胞分离和 FISH 测定显示 PRR34-AS1 的亚细胞定位。RNA 下拉和荧光素酶报告基因测定用于探索 miR-498 是否与 PRR34-AS1、TOMM20 或 ITGA6 结合。进行 Western blot 以检测蛋白质表达。进行挽救实验以验证 PRR34-AS1、miR-498、TOMM20 和 ITGA6 之间的关系。
结果
PRR34-AS1、TOMM20 和 ITGA6 在 HCC 细胞系中表达明显升高，而 miR-498 表达降低。PRR34-AS1、TOMM20 和 ITGA6 促进 HCC 进展，而 miR-498 抑制 HCC 中的细胞增殖、迁移和侵袭。此外，PRR34-AS1、TOMM20 和 ITGA6 与 miR-498 结合。
结论
PRR34-AS1 通过调节 miR-498/TOMM20/ITGA6 轴促进 HCC 进展。
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