摘要
目的检测地西他滨对A431细胞生物学功能的影响并初步探讨其机制。方法将0、5、10和20μmol/L的地西他滨分别作用于A431细胞,通过MTT法观察地西他滨对细胞增殖的影响,通过Western blotting和实时PCR检测地西他滨作用于A431细胞后细胞增殖相关蛋白Cyclin B1、PCNA和CDC2的改变。通过Hoechst 33258染色检测0、5和10μmol/L地西他滨对A431细胞凋亡状态的影响,采用Western blotting和实时PCR检测地西他滨对A431细胞凋亡相关蛋白Bax和Bcl-2的影响。采用transwell实验检测细胞侵袭转移情况,并通过Western blotting和实时PCR检测0、5和10μmol/L的地西他滨对侵袭转移相关蛋白MMP2的影响。结果 MTT检测发现,5、10和20μmol/L的地西他滨作用于A431细胞24 h后对细胞增殖的抑制率分别为(43.81±1.53)%、(48.64±4.65)%和(50.69±4.99)%,随着药物的浓度增加,对A431细胞的增殖抑制率显著增加(P<0.05)。Hoechst 33258染色发现,地西他滨可以显著促进细胞凋亡,transwell实验也证明地西他滨可以明显抑制A431细胞的转移能力。Western blotting和实时PCR结果显示,地西他滨影响A431细胞中Cyclin B1、CDC2、PCNA、Bax、Bcl-2和MMP2蛋白及m RNA水平,地西他滨处理组与对照组比较,差异均有统计学意义(P<0.05)。结论地西他滨对外阴鳞状细胞癌的生物学功能有很强的抑制作用,可以作为治疗外阴鳞状细胞癌的化疗药物。
Objective To investigate the effect of decitabine on the biological function of A431 cells and examine its mechanism. Methods Different concentrations （ 5,10, and 20 μmol/L） of decitabine were used to treat A431 cells. The effect of decitabine on cell proliferation was observed on MTT. Hoechst 33258 staining was used to detect A431 cell apoptosis after the administration of decitabine. The metastasis of A431 cells was detected via transwell assay. The expressions of the proteins related to cell proliferation, apoptosis, invasion, and metastasis were detected using Western blotting and real-time polymerase chain reaction （ PCR）. Results The MTF assay revealed that after treatment with 5,10, and 20μmol/L decitabine, the inhibition rates of cell proliferation were 43.81% ± 1.53%, 48.64% ± 4.65%, and 50.69% ± 4.99%. With the increase in drug concentration, the rate of inhibition of A431 cell proliferation increased significantly （ P 〈 0.05）. Hoechst 33258 staining showed that this treatment could promote apoptosis significantly. The transwell experiments showed that decitabine significantly inhibited the metastasis of A431 cells. Western blotting and real-time PCR showed that the expressions of Cyclin B1, CDC2, Bax, Bcl-2 and MMP2 protein and mRNA in A431 cells were significantly affected by decitabine （P 〈 0.05）. Conclusion Decitabine has a strong inhibitory effect on the biological function of vulvar squamous cell carcinoma and can be used as a chemothera- peutic agent for vulvar squamous cell carcinoma.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2017年第12期1105-1110,共6页
Journal of China Medical University
基金
沈阳市科学技术计划(F15-199-1-34)
关键词
地西他滨
A431
增殖
凋亡
转移
decitabine
A431
proliferation
apoptosis
metastasis
