Effects of FAT10 gene silencing on apoptosis of human esophageal cancer cells and the characteristics of cancer stem cells
10.3969/j.issn.2095-4344.2016.50.001
- VernacularTitle:FAT10基因沉默对人食管癌细胞凋亡及肿瘤干细胞特性的影响
- Author:
Zhiqiang WANG
;
Jianjun ZHANG
- From:
Chinese Journal of Tissue Engineering Research
2016;20(50):7453-7459
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Human leukocyte antigen F-associated transcription factor 10 (FAT10) is highly expressed in many tumor cel s like colon cancer cel s, but its relationship with esophageal cancer is less reported. OBJECTIVE:To investigate the effects of siRNA interference technique on the invasion, apoptosis and the characteristics of EC9706 cel s, a human esophageal cancer cel line. METHODS:siRNA sequence was designed and synthesized according to the FAT10 mRNA encoding sequence, and the EC9706 cel s were transiently transfected. EC9706 cel s were divided into three groups:siRNA FAT10 group, negative control group, and blank control group. The expression levels of bcl-2 and FAT10 were detected by RT-PCR and western blot assay, respectively. Cel counting kit-8 assay was used to measure the proliferation of cel s in vitro. Flow cytometry was used to observe the changes of cel cycle, cel apoptosis and the expression of CD44+CD133+. TUNEL staining was used to detect the apoptosis of the cel s. Cel invasion in vitro was detected by Transwel invasion assay. RESULTS AND CONCLUSION:RT-PCR and western blot findings showed that compared with the negative control group and blank control group, the expression levels of bcl-2 and FAT10 mRNA and protein were significantly decreased in the siRNA FAT10 group (P<0.05);the percentage of CD44+CD133+cel s was decreased significantly (P<0.05);and significantly increased apoptosis rate, and decreased cel proliferation and invasion were also found in the siRNA FAT10 group (P<0.05). In conclusion, the specific silencing of FAT10 gene can reduce the invasion of esophageal cancer cel s, inhibit cel proliferation, reduce bcl-2 expression, and increase the apoptosis rate. Meanwhile, the proportion of CD44+CD133+cel s is decreased.
