Effect of ERH gene knockdown on the proliferation and apoptosis of T24 cells in human bladder cancer
10.3760/cma.j.issn.1006-9801.2018.11.003
- VernacularTitle:抑制未成熟同源蛋白增强子基因的表达对人膀胱癌T24细胞增殖和凋亡的影响
- Author:
Kun PANG
1
;
Lin HAO
;
Zhenduo SHI
;
Bo CHEN
;
Zhiguo ZHANG
;
Rongsheng ZHOU
;
Guanghui ZANG
;
Fei ZHOU
;
Zijian SONG
;
Tian XIA
;
Xitao WANG
;
Zhenning WEI
;
Conghui HAN
Author Information
1. 221009,江苏省徐州市中心医院泌尿外科
- Keywords:
Gene;
ERH;
Urinary bladder neoplasms;
Cell proliferation;
Apoptosis;
Gene knockout;
T24 cell line
- From:
Cancer Research and Clinic
2018;30(11):729-734
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of ERH gene knockdown on the proliferation and apoptosis of human bladder cancer T24 cells. Methods T24 cells infected by lentivirus with interference on ERH gene sequence were cloned to establish stable T24 cells clone in ERH gene suppression. The expression of ERH mRNA gene in bladder cancer was detected by using quantitative real time polymerase chain reaction (qPCR). The effects of ERH knockout on the cell proliferation and apoptosis were examined by using methylthiazolyl tetrazolium (MTT) assay, colony formation assay and flow cytometry. The effect of ERH knockout on the tumorigenic effect of T24 cells in vivo was verified by subcutaneous tumor formation in nude mice. Results After lentiviral transfection, qPCR results showed that the knockdown effect of ERH mRNA in ERH normal group (untreated T24 cells) was better than that in ERH gene knockdown group, and the difference was statistically significant [(1.006±0.126) vs. (0.079±0.007); t=12.72, P=0.0002]. After knocking out ERH gene, MTT assay showed that the proliferation ability of T24 cells in ERH gene knockdown group was weakened compared with ERH normal group, and the difference was statistically significant [A490 value: (0.13±0.00) vs. (0.66±0.01);t=104.61, P<0.0001]. Colony formation assay indicated that the ability of clone in ERH normal group was weakened compared with ERH gene knockdown group [(10.5 ±1.2) vs. (196.4 ±4.0); t= 73.63, P< 0.0001]. Flow cytometry showed that the cell apoptosis rate in ERH gene knockdown group was higher than that in ERH normal group [(11.0 ±0.5) ％ vs. (4.2 ±0.5) ％; t= 16.06, P<0.0001]. Imaging results of subcutaneous tumor formation in nude mice showed that the total fluorescence intensity of the tumor area in ERH gene knockdown group was (4.67 ±0.59) × 1010 μW/cm2, and the corresponding part in ERH normal group was (9.54±4.20) × 1010μW/cm2 (t=3.64, P=0.0051);tumor weight in ERH gene knockdown group was (0.80±0.62) g, and in ERH normal group was (1.79±0.71) g (t=3.33, P=0.0037). Conclusion ERH gene knockout can inhibit the proliferation of human bladder cancer T24 cells, and promote the cell apoptosis.
