c-Myc promoted migration and invasion of malignant melanoma B16-F10 cells through regulating PIK3 R3
10.3969/j.issn.1000-484X.2017.01.013
- VernacularTitle:c-Myc调控PIK3 R3抑制黑色素瘤B16-F10细胞的迁移和侵袭
- Author:
Jianing YANG
;
Fei DENG
;
Ning PAN
- Keywords:
miR-199a;
c-Myc;
Malignant melanoma;
Western blot;
Transwell;
Wound healing assay;
PIK3R3
- From:
Chinese Journal of Immunology
2017;33(1):66-71
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect and the related mechanism of c-Myc on the proliferation,invasion and migration ability of malignant melanoma B16-F10 cells. Methods:We detected the expression of PIK3R3 in malignant melanoma and normal tis-sues. Efficiency of gene silencing of c-Myc and PIK3R3 was determined by qPCR and Western blot. We detected the proliferate ability of B16-F10 cells after silencing c-Myc and PIK3R3 using EdU assay. We detected the migration and invasion ability of B16-F10 cells after silencing c-Myc and PIK3R3 using wound healing assays and Transwell matrigel invasion assays. The expression of miR-199a after silencing c-Myc and PIK3R3 using qPCR. The expression of c-Myc and PIK3R3 was detected by qPCR after transfecting miR-199a mimics or miR-199a inhibitor. Dual-luciferase reporter assay system was used to detect miR-199a regulating c-Myc and PIK3R3. Results:Compared normal skin tissue,expression of PIK3R3 was significantly increased in malignant melanoma tissue;after silencing c-Myc and PIK3R3 gene,the proliferation,invasion and metastasis of melanoma cell line B16-F10 were significantly reduced;expression of miR-199a upregulated after silencing c-Myc and PIK3R3 genes, expression of c-Myc and PIK3R3 decreased after transfecting miR-199a mimics, expression of c-Myc and PIK3R3 upregulated after transfecting miR-199a inhibitor, dual luciferase reporter system test results revealed miR-199a can directly regulate c-Myc and PIK3R3 transcription activity. Conclusion: miR199a regulated the expression of c-Myc,then promote proliferation,migration and invasion in malignant melanoma cells.
