LncRNA-PVT1/miR-15a/Bmi-1 Pathway Regulates the Proliferation of HGC-27 Gastric Cancer Cells in Vitro
10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0508
- VernacularTitle:LncRNA-PVT1/miR-15a/Bmi-1通路调控HGC-27胃癌细胞的体外增殖
- Author:
Jing-ying HOU
1
;
Ya-xian WU
2
;
Xiao-yan JIN
3
;
Hui LING
4
;
Ting-feng YU
2
;
Ling-yun WANG
2
Author Information
1. Clinical Research Center， Sun Yat-sen Memorial Hospital of Sun Yat-sen University， Guangzhou 510120， China
2. Department of Gastroenterology， Sun Yat-sen Memorial Hospital of Sun Yat-sen University， Guangzhou 510120， China
3. General Department， Sun Yat-sen Memorial Hospital of Sun Yat-sen University， Guangzhou 510120， China
4. Shenzhen People's Hospital，Shenzhen 518020， China
- Publication Type:Journal Article
- Keywords:
lncRNA-PVT1;
miR-15a;
Bmi-1;
gastric cancer cell;
proliferation
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2021;42(5):703-713
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the role of lncRNA-PVT1/miR-15a/ Bmi-1 pathway in the proliferation of HGC-27 gastric cancer cells in vitro. MethodsGastric cancer cell line HGC-27 was cultured in vitro and divided into the following groups： （1） blank control， si-PVT1 and si-PVT1 NC. The si-PVT1 and si-PVT1 NC groups were transferred with PVT1 siRNA and PVT1 siRNA scramble， respectively. （2） blank control， miR-15a and miR-15a NC. （3） blank control， si-Bmi-1 and si-Bmi-1 NC. Cell proliferation was evaluated by using MTS. The expression of PVT1， miR-15a and Bmi-1 was detected in the condition of PVT1 inhibition. In order to validate the relationship between miR-15a and Bmi-1， miR-15a mimic was transfected into HGC-27 gastric cancer cell line， and the expression of miR-15a and Bmi-1 was examined. The potential complementary binding sites of PVT1 and miR-15a， and miR-15a and Bmi-1 were predicted by bioinformatics. Dual-luciferase reporter assay was performed to detect luciferase activity of the different groups of cells in order to verify the relationship between PVT1 and miR-15a and miR-15a and Bmi-1. Relevant rescue experiment was conducted in order to further validate the mutual relationship of lncRNA-PVT1， miR-15a and Bmi-1. ResultsOD490 value and cell proliferation rate were significantly decreased in the condition of PVT1 or Bmi-1 inhibition， or miR-15a overexpression （P<0.01）. The expression of Bmi-1 was decreased in si-PVT1 group， whereas miR-15a was increased （P<0.01）. miR-15a was increased after transfection， whereas the expression of Bmi-1 was decreased （P<0.01）. The dual luciferase reporter assay indicated that both the PVT1 and Bmi-1 reporter gene luciferase activity were decreased significantly in miR-15a mimics group， down-regulating 56% and 32%， respectively （P<0.01）. The expression of PVT1 and Bmi-1 was elevated in the condition of PVT1 knockdown and miR-15a inhibition， whereas both of their expression were further decreased in the condition of PVT1 knockdown and miR-15a transfection. ConclusionLncRNA-PVT1 could upregulate Bmi-1 （lncRNA-PVT1/miR-15a/Bmi-1 pathway） to promote the proliferation of gastric cancer cells in vitro by inhibiting miR-15a.
