摘要
目的探讨着色性干皮病基因C(xeroderma pigmentosum group C,XPC)对膀胱癌T24细胞紫外线(ultraviolet ray,UV)介导的DNA损伤修复的作用及机制。方法利用shRNA策略,Western blot鉴定XPC沉默效果,建立稳定抑制XPC的膀胱肿瘤T24细胞模型;CCK-8法检测UV对细胞增殖的影响;宿主反应实验检测细胞DNA损伤修复能力的差异;免疫荧光检测UV损伤处理、XPC干扰前后p-ATM蛋白分子向细胞核DNA损伤部位募集情况。结果成功建立稳定抑制XPC的膀胱肿瘤T24细胞模型,干扰XPC后,细胞对UV损伤敏感性出现显著差异(P<0.01),干扰组DNA损伤修复能力较对照组降低(P<0.01),p-ATM向损伤区域的募集作用降低(P<0.01)。结论 UV损伤条件下,膀胱癌T24细胞通过XPC介导p-ATM向损伤区域的募集,以降低细胞UV损伤修复能力。
Objective To determine the role and the underlying mechanisms of xeroderma pigmentosum group C (XPC) in ultraviolet (UV) DNA damage response of human bladder cancer cell line T24. Methods XPC siRNA was used to induce T24 cells with stable XPC knockdown, and the obtained cells were confirmed by Western blotting. CCK-8 kit was employed to detect the inhibitory effects of UV on the T24 cells and T24 cells with XPC knockdown. The ability of DNA repair was evaluated by host-cell reactivation (HCR) assay. The nuclear recruitment of p-ATM towards the damaged DNA was determined by immunofluoresence staining. Results The T24 cells with stable XPC knockdown were successfully established. Compared to the normal T24 cells, the XPC knockdown cells were more sensitive to UV treatment (P〈0.01), showed declined ability of DNA repair (P〈0.01), and reduced nuclear concentration of p-ATM towards the damaged DNA(P〈0.01). Conclusion XPC play a major role in DNA repair via affecting the nuclear recruitment of p-ATM in T24 cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2014年第9期856-859,共4页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(30972979)~~