Acquisition of multidrug resistance by overexpression of Twist1 in colorectal cancer cells SW620 and its mechanism
10.3969/j.issn.1000-484X.2019.03.011
- VernacularTitle:过表达Twist1促进结直肠癌SW620细胞获得多药耐药性及机制研究
- Author:
Jun-Min ZHOU
1
;
Shui-Qing HU
;
Kun-Ming WEN
Author Information
1. 遵义医学院附属医院胃肠外科
- Keywords:
Twist1;
Colorectal cancer;
Multidrug resistance;
Cancer stem cells
- From:
Chinese Journal of Immunology
2019;35(3):306-310
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of overexpression of Twist1 on multidrug resistance( MDR) in colorectal cancer SW620 cells and its possible mechanism. Methods: The colorectal cancer SW620 cells were transfected with Twist1 overexpressing lentivirus and negative control virus, which were called experimental group ( SW620-Twist1) and control group ( SW620-NC) respectively. The protein and mRNA expression of Twist1、ABCB1 and ABCG2 in two groups were detected by Western blot and RT-qPCR, respectively. The growth inhibitory rate of 5-Fluorouracil(5-FU) and Oxaliplatin( OXA) at 24 ,48 and 72 h in two groups were determined by CCK8 assay. Cancer stem cell markers CD133,CD44 expression levels were detected by Flow cytometry. Results: The experimental group was compared with the control group, the expressions of Twsit1 gene mRNA and protein increased significantly( P<0. 01) ,Twist1 gene overexpressing SW620 cell line was successfully constructed. The inhibition rates of cell proliferation of the three chemotherapeutic drugs at 48 h and 72 h were significantly decreased ( 48 h, P<0. 05 ; 72 h, P<0. 01) . The mRNA and protein expression levels of ABCB1,ABCG2 were significantly increased( P<0. 01). The expression of CSCs markers CD133 and CD44 increased significantly (P<0. 01). Conclusion: Overexpression of Twist1 can promote the expression of ABCB1 and ABCG2 genes in SW620 cells and make the tumor cells acquire MDR. This phenomenon may be related to the promotion of stemness.
