中国实验诊断学
中國實驗診斷學
중국실험진단학
Chinese Journal of Laboratory Diagnosis
2015年
10期
1632-1635
,共4页
自噬%PI3K/mTOR 抑制剂 NVP-BEZ235%3-MA%凋亡%人结肠癌
自噬%PI3K/mTOR 抑製劑 NVP-BEZ235%3-MA%凋亡%人結腸癌
자서%PI3K/mTOR 억제제 NVP-BEZ235%3-MA%조망%인결장암
autophagy%dual PI3K/mTOR inhibitor NVP-BEZ235%3-MA%Apoptosis%human colon adenocarcinoma cell
目的：探讨自噬对 I3K／mTOR 抑制剂 NVP-BEZ235诱导的人结肠腺癌 DLD1细胞凋亡的影响。方法PI3K／mTOR 抑制剂 NVP-BEZ235和（或）自噬抑制剂3-MA 处理人结肠腺癌 DLD1细胞，MTT 法检测细胞生存率，流式细胞术检测细胞凋亡率的变化，Western Blot 检测凋亡相关蛋白的表达。结果不同浓度的 NVP-BEZ235作用于人结肠腺癌 DLD1细胞24 h 后，MTT 及流式细胞术结果显示，明显地抑制了细胞增殖，并增加了细胞凋亡率，同时观测到凋亡相关蛋白 Caspase-3表达升高；采用3-MA 特异性抑制自噬活性后，明显地增加了 NVP-BEZ235诱导的细胞凋亡率；同时，检测到凋亡相关蛋白 Caspase-3的表达上调。结论自噬在 NVP-BEZ235诱导的人结肠腺癌 DLD1细胞凋亡的过程中起保护作用，3-MA 抑制自噬后，促进了 NVP-BEZ235诱导人结肠腺癌 DLD1细胞凋亡，联合应用PI3K／mTOR 抑制剂 NVP-BEZ235和自噬抑制剂3-MA 有望成为人结肠癌的新治疗策略。
目的：探討自噬對 I3K／mTOR 抑製劑 NVP-BEZ235誘導的人結腸腺癌 DLD1細胞凋亡的影響。方法PI3K／mTOR 抑製劑 NVP-BEZ235和（或）自噬抑製劑3-MA 處理人結腸腺癌 DLD1細胞，MTT 法檢測細胞生存率，流式細胞術檢測細胞凋亡率的變化，Western Blot 檢測凋亡相關蛋白的錶達。結果不同濃度的 NVP-BEZ235作用于人結腸腺癌 DLD1細胞24 h 後，MTT 及流式細胞術結果顯示，明顯地抑製瞭細胞增殖，併增加瞭細胞凋亡率，同時觀測到凋亡相關蛋白 Caspase-3錶達升高；採用3-MA 特異性抑製自噬活性後，明顯地增加瞭 NVP-BEZ235誘導的細胞凋亡率；同時，檢測到凋亡相關蛋白 Caspase-3的錶達上調。結論自噬在 NVP-BEZ235誘導的人結腸腺癌 DLD1細胞凋亡的過程中起保護作用，3-MA 抑製自噬後，促進瞭 NVP-BEZ235誘導人結腸腺癌 DLD1細胞凋亡，聯閤應用PI3K／mTOR 抑製劑 NVP-BEZ235和自噬抑製劑3-MA 有望成為人結腸癌的新治療策略。
목적：탐토자서대 I3K／mTOR 억제제 NVP-BEZ235유도적인결장선암 DLD1세포조망적영향。방법PI3K／mTOR 억제제 NVP-BEZ235화（혹）자서억제제3-MA 처리인결장선암 DLD1세포，MTT 법검측세포생존솔，류식세포술검측세포조망솔적변화，Western Blot 검측조망상관단백적표체。결과불동농도적 NVP-BEZ235작용우인결장선암 DLD1세포24 h 후，MTT 급류식세포술결과현시，명현지억제료세포증식，병증가료세포조망솔，동시관측도조망상관단백 Caspase-3표체승고；채용3-MA 특이성억제자서활성후，명현지증가료 NVP-BEZ235유도적세포조망솔；동시，검측도조망상관단백 Caspase-3적표체상조。결론자서재 NVP-BEZ235유도적인결장선암 DLD1세포조망적과정중기보호작용，3-MA 억제자서후，촉진료 NVP-BEZ235유도인결장선암 DLD1세포조망，연합응용PI3K／mTOR 억제제 NVP-BEZ235화자서억제제3-MA 유망성위인결장암적신치료책략。
Objective To investigate effects of inhibition of autophagy on NVP-BEZ235-induced apoptosis in human colon adenocarcinoma cell line DLD1.Methods After the DLD1 cells were treated with NVP-BEZ235 and/or 3-MA, cell viability was detected by an MTT assay.The rate of cell apoptosis was detected using flow cytometry.The expres-sion of apoptosis-related protein were detected by Western blot.Results The cells were treated with varied concentra-tion of NVP-BEZ235 for 24 h.According to the results of MTT and flow cytometry,the rate of cell proliferation de-creased,and the rate of cell apoptosis increased in the dose of NVP-BEZ235.Meanwhile,the expression of apoptosis-re-lated protein,cleaved caspase 3,was increased.However,after autophagy was inhibited by 3-MA,the rate of cell viabili-ty was obviously decreased.Meanwhlie,we detected that inhibition of autophagy enhanced the rate of NVP-BEZ235-in-duced apoptosis and the expression of apoptosis-related protein were obviously increased.Conclusion Autophagy pro-tected human colon adenocarcinoma cell line DLD1 from NVP-BEZ235-induced apoptosis.In addition,the blockage of autophagy could enhance NVP-BEZ235-induced apoptosis.The combined therapy of NVP-BEZ235 and 3-MA may be a promising therapeutic strategy for human colon adenocarcinoma cell line DLD1.
