中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
6期
865-867
,共3页
陈先国%庄乾元%梁朝朝%杜立环%叶章群
陳先國%莊乾元%樑朝朝%杜立環%葉章群
진선국%장건원%량조조%두립배%협장군
mTORC1%mTORC2%AR%前列腺癌%Akt
mTORC1%mTORC2%AR%前列腺癌%Akt
mTORC1%mTORC2%AR%전렬선암%Akt
mTORC1%mTORC2%AR%Prostate carcinoma%Akt
目的 观察mTORC1和mTORC2在前列腺癌C4-2细胞中的作用.方法 噻唑蓝(MTY)比色法检测转染siRNA raptor和siRNA rictor后C4-2细胞增殖改变;流式细胞术(FCM)检测敲除mTORC1(raptor)和mTORC2(rictor)后C4-2细胞凋亡;Western blot检测siRNA raptor和siRNArictor后C4-2细胞雄激素受体(AR)和Akt磷酸化表达.结果 MTT显示敲除raptor生长抑制率无显著变化[(25.37±2.63)%比(27.49±2.96)%,P＞0.05],而敲除rictor组[(25.37±2.63)%比(62.86±5.61)%,P＜0.01]显著地抑制了细胞的生长;FCM显示敲除raptor显著增加了细胞凋亡[(11.76±1.45)%比(38.23±3.71)%,P＜0.01],而敲除rictor对C4-2细胞凋亡无显著性变化[(11.76±1.45)%比(14.25 ±1.68)%,P＞0.05];Western blot检测显示敲除mTORC1显著增加C4-2细胞AR[(0.21±0.04)%比(0.73 ±0.12)%,P＜0.01]和Akt磷酸化表达[(0.23±0.06)%比(0.68±0.11)%,P＜0.01],而敲除mTORC2显著地抑制了C4-2细胞AR[(0.21 ±0.04)%比(0.07 ±0.02)%,P＜0.01]和Akt磷酸化表达[(0.23±0.06)%比(0.06±0.03)%,P＜0.01].结论 mTORC2对前列腺癌C4-2细胞的存活是必须的,mTORC2是治疗前列腺癌有希望的靶目标.
目的 觀察mTORC1和mTORC2在前列腺癌C4-2細胞中的作用.方法 噻唑藍(MTY)比色法檢測轉染siRNA raptor和siRNA rictor後C4-2細胞增殖改變;流式細胞術(FCM)檢測敲除mTORC1(raptor)和mTORC2(rictor)後C4-2細胞凋亡;Western blot檢測siRNA raptor和siRNArictor後C4-2細胞雄激素受體(AR)和Akt燐痠化錶達.結果 MTT顯示敲除raptor生長抑製率無顯著變化[(25.37±2.63)%比(27.49±2.96)%,P＞0.05],而敲除rictor組[(25.37±2.63)%比(62.86±5.61)%,P＜0.01]顯著地抑製瞭細胞的生長;FCM顯示敲除raptor顯著增加瞭細胞凋亡[(11.76±1.45)%比(38.23±3.71)%,P＜0.01],而敲除rictor對C4-2細胞凋亡無顯著性變化[(11.76±1.45)%比(14.25 ±1.68)%,P＞0.05];Western blot檢測顯示敲除mTORC1顯著增加C4-2細胞AR[(0.21±0.04)%比(0.73 ±0.12)%,P＜0.01]和Akt燐痠化錶達[(0.23±0.06)%比(0.68±0.11)%,P＜0.01],而敲除mTORC2顯著地抑製瞭C4-2細胞AR[(0.21 ±0.04)%比(0.07 ±0.02)%,P＜0.01]和Akt燐痠化錶達[(0.23±0.06)%比(0.06±0.03)%,P＜0.01].結論 mTORC2對前列腺癌C4-2細胞的存活是必鬚的,mTORC2是治療前列腺癌有希望的靶目標.
목적 관찰mTORC1화mTORC2재전렬선암C4-2세포중적작용.방법 새서람(MTY)비색법검측전염siRNA raptor화siRNA rictor후C4-2세포증식개변;류식세포술(FCM)검측고제mTORC1(raptor)화mTORC2(rictor)후C4-2세포조망;Western blot검측siRNA raptor화siRNArictor후C4-2세포웅격소수체(AR)화Akt린산화표체.결과 MTT현시고제raptor생장억제솔무현저변화[(25.37±2.63)%비(27.49±2.96)%,P＞0.05],이고제rictor조[(25.37±2.63)%비(62.86±5.61)%,P＜0.01]현저지억제료세포적생장;FCM현시고제raptor현저증가료세포조망[(11.76±1.45)%비(38.23±3.71)%,P＜0.01],이고제rictor대C4-2세포조망무현저성변화[(11.76±1.45)%비(14.25 ±1.68)%,P＞0.05];Western blot검측현시고제mTORC1현저증가C4-2세포AR[(0.21±0.04)%비(0.73 ±0.12)%,P＜0.01]화Akt린산화표체[(0.23±0.06)%비(0.68±0.11)%,P＜0.01],이고제mTORC2현저지억제료C4-2세포AR[(0.21 ±0.04)%비(0.07 ±0.02)%,P＜0.01]화Akt린산화표체[(0.23±0.06)%비(0.06±0.03)%,P＜0.01].결론 mTORC2대전렬선암C4-2세포적존활시필수적,mTORC2시치료전렬선암유희망적파목표.
Objective To investigate the role of mTORC1 and mTORC2 in prostate cancer C4-2 cells. Methods The growth inhibition and apoptosis rate were examined by methyl thiazol tetrazolium ( MTT) assay and flow cytometry ( FCM) after knockouting raptor and rictor in prostate cancer C4-2 cells.The expression of androgen receptor ( AR) and Akt phosphorylation after transfection of siRNA raptor and rictor was detected by Western blotting. Results The growth inhibition of C4-2 cells had no significant change after transfecting siRNA raptor [(25. 37 ± 2. 63) % vs (27.49 ± 2. 96) % , P ＞ 0.05] , and the apoptosis rate was markedly increased [(11. 76 ± 1. 45) % vs (38. 23 ± 3. 71) % ,P ＜0. 01]. The inhibition of mTORC2 (rictor) markedly decreased the growth of C4-2 cells [(25.37 ±2.63)% vs (62.86 ±5.61)% ,P＜0.01] , and the apoptosis rate had no significant change [(11.76 ±1.45)% vs (14.25±1.68)%,P＞0.05]. The expression of AR [(0.21 ±0.04)% vs (0. 73 ±0. 12)% ,P＜0. 01] and Akt phosphorylation [(0. 23 ± 0. 06 ) % vs ( 0. 68 ± 0. 11 ) % , P ＜ 0. 01] were significantly increased after knocking down mTORC1 (raptor) in C4-2 cells, andt the inhibition of mTORC2 (rictor) markedly decreased the expression of AR [( 0. 21 ± 0. 04 ) % vs ( 0. 07 ± 0. 02 ) % , P ＜ 0. 01] and Akt phosphorylation [(0. 23 ± 0. 06) % vs ( 0. 06 ± 0. 03) % , P ＜ 0. 01]. Conclusion mTORC2 not only is required for the survival of prostate cancer, but also a promising therapic target.
