miR-338-3p Targets and Regulates RNF121 to Promote the Proliferation and Invasion of Human Lung Cancer A549 Cells
10.13865/j.cnki.cjbmb.2021.04.1002
- Author:
Jing-Chen XIE
1
;
Chuan LI
1
;
Zhan-Qing ZHAO
1
;
Xie-Ju XIE
2
Author Information
1. Department of Respiratory Medicine, Hainan Western Central Hospital
2. Department of Pathophysiology, Hainan Medical College
- Publication Type:Journal Article
- Keywords:
non-small cell lung cancer;
normal lung cancer A549;
ring finger protein 121(RNF121);
tumor formation in nude mice
- From:
Chinese Journal of Biochemistry and Molecular Biology
2021;37(7):908-916
- CountryChina
- Language:Chinese
-
Abstract:
Recent studies have shown that miR-338-3p plays an important role in the proliferation and invasion of lung cancer, but whether miR-338-3p regulates lung cancer proliferation and invasion through targeting ring finger protein 121 (RNF121) is still unclear. In order to explore its mechanism, the normal lung cell line MRC-5 and the non-small cell lung cancer line A549 were cultured in vitro. Using qRTPCR and Western blotting detection, we found that the expression of miR-338-3p in A549 lung cancer cells was lower than that in MRC-5 cells, while RNF121 expression increased (P<0. 05). The miR-338-3p bound to the 3′UTR of RNF121 was predicted by TargetScan (http:/ / www. targetscan. org/mamm_31/) target gene prediction software. The dual luciferase reporter confirmed the complementary combination. We constructed a shRNA vector targeting RNF121 and transfected cells, used CCK8 experiments to detect cell proliferation in each group, employed Annexin V-FITC/PI double staining to detect cell apoptosis, performed Transwell experiments and cell colony formation experiments to detect cell invasion and clonal proliferation ability of each group. Cells of each group were inoculated under the skin of nude mice and we drew the tumor growth curve and the size of the tumor were measured. Then the tumor tissue was obtained and TUNEL staining was used to detect apoptosis in the tissue. Compared with the blank group, the cell proliferation, invasion, and cell cloning ability in the miR-338-3p mimic and sh-RNF121 groups were inhibited, but apoptosis was significantly induced. At the same time, in vivo experiments found that the ability to form tumors was reduced, and apoptosis in tumor tissues was increased (P<0. 05). The miR-338-3p group had a higher cell proliferation activity, invasion and colony-forming ability but lower tumorigenicity and apoptosis in tumor tissues (P<0. 05). Compared with the blank group, no significant difference was found in the NC group and miR-338-3p inhibitor+sh-RNF121 group (P>0. 05). In summary, miR-338-3p can target the expression of RNF121 to inhibit the proliferation and invasion of A549 cells and inhibit the growth of subcutaneous transplanted tumors in nude mice. RNF121 is expected to become a new target for the treatment of non-small cell lung cancer.
