KLF4沉默与阿霉素诱导的DNA损伤协同抑制肝癌HepG2细胞的生长

KLF4 silencing and doxorubicin-induced DNA damage inhibit the growth of hepatocellular carcinoma HepG2 cells

探究KLF4沉默与经不同作用浓度阿霉素处理诱导的DNA损伤对肝癌HepG2细胞增殖凋亡的影响及其作用机制.应用RNA干扰技术,采用siRNA转染HepG2细胞以沉默KLF4基因.采用MTT法检测KI.F4沉默前后对HepG2细胞增殖的影响,使用流式细胞术检测KLF4沉默前后对HepG2细胞周期变化影响,应用Western blot法检测转染前后HepG2细胞中KLF4蛋白及细胞周期相关蛋白表达变化.Western blot检测到高浓度的阿霉素促进KLF4的表达,并且低浓度的阿霉素可使得细胞停滞在Gz/M期,高浓度的阿霉素则使部分细胞凋亡(19.31%).将KLF4沉默后,发现细胞生长变缓,低浓度的阿霉素处理后,细胞随时间增加而出现更多的细胞凋亡;高浓度的阿霉素处理后,细胞数明显减少,更多的细胞发生凋亡(2& 89%),且在KLF4沉默前后均发现低浓度阿霉素促进p53与p21表达,高浓度阿霉素抑制其表达.阿霉素诱导的DNA损伤可提高KLF4的表达,KLF4依赖于DNA损伤激活的p53促进p21的表达,进而引起G〃S期细胞周期阻滞.沉默KLF4与阿霉素诱导的DNA损伤可协同抑制肝癌细胞的增殖、促进凋亡,其在肝癌细胞HepG2中扮演十分重要的角色.

In this paper, the effect of KLF4 silencing and DNA damage induced by different concentrations of doxorubicin on proliferation and apoptosis of HepG2 cells and its related mechanism were investigated. RNA interference technique was applied to siRNA transfection of HepG2 cells to silence KLF4 genes. And the effect of KLF4 on HepG2 cell proliferation treated with cytostatic concentration and cytotoxic concentration on doxorubicin were detected by MTT assay before and after KLF4investigated on top of that, so as to provide reference for the prevention and control of pollution regionally. The results show as follows. 1 ) The concentration of 60 monitoring sites differ a lot and the average PM2.s concentration of sites exceeds the second level of the national ambient air quality standard. The concentration is significantly affected by four factors, namely temperature, wind velocity, altitude and area of green space in buffer with 3 km radius? The rise of temperature will be contributed to increase the value of PM2.5? while other factors are opposite? 2) The spatial distribution simulation of PM2 5 based on LUR shows obvious gradient differences of PM2.5 concentration, which are high in the middle region, low in the east, and the lowest in the west on a provincial scale? Generally, the pollution in Wuhan Metropolitan, Jianghan Plain and part of Xiangyang are more severe than the western parts relatively. 3) Taking population density into consideration, the study of PM2<5 exposure intensity index built exhibits a high spatial correlation between the exposure intensity and population density. The majority of population and land in the province is in low exposure intensity. The highly exposed areas are comparatively dispersed, mainly distributed in the middle and east of Hubei centered in Wuhan metropolitan.