中国临床医学
中國臨床醫學
중국림상의학
CLINICAL MEDICAL JOURNAL OF CHINA
2015年
2期
143-146
,共4页
结肠癌%OSI-027%哺乳动物雷帕霉素靶蛋白%凋亡%信号转导
結腸癌%OSI-027%哺乳動物雷帕黴素靶蛋白%凋亡%信號轉導
결장암%OSI-027%포유동물뢰파매소파단백%조망%신호전도
Colon cancer%OSI-027%Mammalian target of rapamycin%Apoptosis%Signal transduction
目的：探讨新型mTORC1／mTORC2双重抑制剂OSI‐027对人结肠癌细胞株HT‐29的体外抑制作用。方法：体外培养的人结肠癌HT‐29细胞株，给予不同浓度（0．1、1、10、25、50 nmol／L）的OSI‐027处理，或以25 nmol／L OSI‐027处理0、24、48、72、96 h后，采用四甲基偶氮唑盐（methyl thiazolyl tetrazolium ，MTT）法、细胞集落形成实验观察HT‐29细胞的生长增殖情况；应用流式细胞仪（FACS）及台盼蓝染色法检测OSI‐027处理后 HT‐29细胞的凋亡和死亡情况；采用蛋白质印迹（Western blotting）法检测HT‐29细胞凋亡相关蛋白cleaved‐caspase‐3和细胞色素C（cytochrome C）的表达。结果：OSI‐027可抑制HT‐29细胞的存活，且呈浓度和时间依赖性；还可抑制HT‐29细胞的增殖，呈浓度依赖性；此外，OSI‐027还能诱导HT‐29细胞的凋亡和死亡，并呈浓度依赖性。OSI‐027处理后，HT‐29细胞中凋亡相关蛋白cleaved‐caspase‐3和cytochrome C的表达水平上调。结论：新型mTORC1／mTORC2双重抑制剂OSI‐027可抑制人结肠癌细胞HT‐29的增殖并诱导细胞凋亡。
目的：探討新型mTORC1／mTORC2雙重抑製劑OSI‐027對人結腸癌細胞株HT‐29的體外抑製作用。方法：體外培養的人結腸癌HT‐29細胞株，給予不同濃度（0．1、1、10、25、50 nmol／L）的OSI‐027處理，或以25 nmol／L OSI‐027處理0、24、48、72、96 h後，採用四甲基偶氮唑鹽（methyl thiazolyl tetrazolium ，MTT）法、細胞集落形成實驗觀察HT‐29細胞的生長增殖情況；應用流式細胞儀（FACS）及檯盼藍染色法檢測OSI‐027處理後 HT‐29細胞的凋亡和死亡情況；採用蛋白質印跡（Western blotting）法檢測HT‐29細胞凋亡相關蛋白cleaved‐caspase‐3和細胞色素C（cytochrome C）的錶達。結果：OSI‐027可抑製HT‐29細胞的存活，且呈濃度和時間依賴性；還可抑製HT‐29細胞的增殖，呈濃度依賴性；此外，OSI‐027還能誘導HT‐29細胞的凋亡和死亡，併呈濃度依賴性。OSI‐027處理後，HT‐29細胞中凋亡相關蛋白cleaved‐caspase‐3和cytochrome C的錶達水平上調。結論：新型mTORC1／mTORC2雙重抑製劑OSI‐027可抑製人結腸癌細胞HT‐29的增殖併誘導細胞凋亡。
목적：탐토신형mTORC1／mTORC2쌍중억제제OSI‐027대인결장암세포주HT‐29적체외억제작용。방법：체외배양적인결장암HT‐29세포주，급여불동농도（0．1、1、10、25、50 nmol／L）적OSI‐027처리，혹이25 nmol／L OSI‐027처리0、24、48、72、96 h후，채용사갑기우담서염（methyl thiazolyl tetrazolium ，MTT）법、세포집락형성실험관찰HT‐29세포적생장증식정황；응용류식세포의（FACS）급태반람염색법검측OSI‐027처리후 HT‐29세포적조망화사망정황；채용단백질인적（Western blotting）법검측HT‐29세포조망상관단백cleaved‐caspase‐3화세포색소C（cytochrome C）적표체。결과：OSI‐027가억제HT‐29세포적존활，차정농도화시간의뢰성；환가억제HT‐29세포적증식，정농도의뢰성；차외，OSI‐027환능유도HT‐29세포적조망화사망，병정농도의뢰성。OSI‐027처리후，HT‐29세포중조망상관단백cleaved‐caspase‐3화cytochrome C적표체수평상조。결론：신형mTORC1／mTORC2쌍중억제제OSI‐027가억제인결장암세포HT‐29적증식병유도세포조망。
Objective:To investigate the inhibitory effect of novel mTORC1/mTORC2 dual inhibitor OSI‐027 on human colon cancer cell line HT‐29 in vitro .Methods:HT‐29 cells ,cultured in vitro ,were either treated with different concentrations of OSI‐027(0 .1 ,1 ,10 ,25 and 50 nmol/L) ,or treated with 25 nmol/L OSI‐027 for 0 ,24 ,48 ,72 ,96 h ,respectively .Methyl thiazolyl tetrazolium assay(MTT) and cell colony forming assay were used to detect the growth and proliferation of HT‐29 cells after treatment with OSI‐027 .Flow cytometry (FACS) and trypan blue stalning were used to detect the influence of OSI‐027 on apoptosis and death of HT‐29 cells .Western blotting was used to detect the expression of apoptosis‐related proteins cleaved‐caspase‐3 and cytochrome C .Results:OSI‐027 inhibited the survival of HT‐29 cells ,and the inhibition was concentration and time dependent .It also inhibited the proliferation of HT‐29 cells ,and the inhibition was concentration dependent .It also induced apoptosis and death ,and the inducement was concentration dependent .Expression levels of apoptosis‐related proteins cleaved‐caspase‐3 and cytochrome C increased after OSI‐027 treatment .Conclusions:The novel mTORC1/mTORC2 dual inhibitor OSI‐027 can inhibit the proliferation of human colon cancer cell HT‐29 and induce cell apoptosis .
