miR-222 promotes retinoblastoma cell proliferation and invasion by targeting RB1
10.19401/j.cnki.1007-3639.2016.09.004
- VernacularTitle:miR-222通过靶向RB1促进视网膜母细胞瘤细胞生长与侵袭
- Author:
Yuefeng LIU
;
Yong ZHANG
;
Xiaodong ZHONG
;
Weimin LUO
- Publication Type:Journal Article
- Keywords:
Retinoblastoma;
miR-222;
RB1;
Growth;
Invasion
- From:
China Oncology
2016;26(9):743-749
- CountryChina
- Language:Chinese
-
Abstract:
Background and purpose:A large number of studies have showed that retinoblastoma gene 1 (RB1) can inhibit the occurrence and development of many tumors, including neuroblastoma, small cell lung cancer, osteosar-coma, pancreatic cancer, breast cancer and so on. RB1 is also closely related to the regulation of cell cycle, differentia-tion, senescence, apoptosis, growth inhibition, etc. The goal of this article is to elucidate whether miR-222 promotes cell proliferation and invasion by targeting RB1, further to explore the molecular mechanism that miR-222 functions as an oncogene in retinoblastoma cells.Methods:miR-222 (miR-222 mimics) and RB1-wt, miR-NC and RB1-wt, miR-222 and RB1-mut, miR-NC (a controlled miR-222 mimics) and RB1-mut were co-transfected into Y79 cells, and luciferase activity was detected by single photon. Retinoblastoma cells were transfected with miR-222 mimics and miR-NC, and the expressions of RB1 protein were detected by Western blot. Retinoblastoma cell proliferation assays were performed by MTS assay when miR-222, miR-NC, RB1 (pcDNA3.1-RB1), vector (pcDNA3.1), miR-222+RB1 and miR-NC+vec-tor were transfected into Y79 cells. The growth and invasion ability of Y79 cells with ectopic expression of miR-222 were evaluated by MTS and Transwell invasion assays.Results:This study demonstrated that miR-222 could promote the luciferase activity of RB1-wt. The expression levels of luciferase reporter gene activity in Y79 cells after transfection with miR-222+RB1-wt were higher than those in the negative control cells (miR-NC+RB1-wt) (P<0.05). The protein expression levels of RB1 in Y79 cells after transfection with miR-222 were lower than those in miR-NC (P<0.05). Overexpression of RB1 inhibited the proliferation of retinoblastoma cells. miR-222 promoted the prolifera-tion of retinoblastoma cells through targeting RB1 (P<0.05). Moreover, there was no signiifcant difference between the cell survival rates of Y79 which were transfected with miR-222+pcDNA3.1-RB1 and miR-NC+pcDNA3.1 (P>0.05). After transfection with miR-222 mimics for 48 h, Transwell invasion assay showed that the number of cells through the basement membrane was (193±10). Compared with the control group (144±11), it could signiifcantly accelerate the invasion of Y79 cells (P<0.01). There was no signiifcant difference between the number of cells through the basement membrane which were transfected with miR-222+pcDNA3.1-RB1 and miR-NC+pcDNA3.1 (P>0.05).Conclusion:miR-222 promotes cell proliferation and invasion by targeting RB1 expression in retinoblastoma cells.
