中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2008年
34期
2427-2431
,共5页
姜淑娟%张嵩%牟晓燕%李洧%王艳
薑淑娟%張嵩%牟曉燕%李洧%王豔
강숙연%장숭%모효연%리유%왕염
子宫内膜肿瘤%凋亡%微管蛋白%紫杉醇%曲古抑菌素A
子宮內膜腫瘤%凋亡%微管蛋白%紫杉醇%麯古抑菌素A
자궁내막종류%조망%미관단백%자삼순%곡고억균소A
Endometrial neoplasms%Aoptposis%Microtubulin%Paclitaxel%Trichostetin A
目的 探讨曲古抑菌素A(TSA)和紫杉醇(PTX)体外对人子宫内膜癌细胞凋亡和微管稳定性的影响及其机制.方法 将浆液性乳头状子宫内膜癌Ark2细胞、低分化子宫内膜样腺癌KLE和AN3细胞各分为TSA、PTX单独作用组(TSA组、PTX组)和联合作用组(TSA+PTX组),并设空白对照组,以锥虫蓝拒染法观察药物对细胞增殖的影响,膜联蛋白V、Hoechst染色法检测细胞凋亡率,流式细胞仪测定线粒体膜电位(MMP),蛋白质印迹法检测半胱天冬酶9(caspase-9)、多聚ADP核糖聚合酶(PARP)和乙酰化微管蛋白的表达.结果 TSA、PTX对Ark2、KLE和AN3细胞均有抑制作用,二药联用后抑制作用更强.药物作用后4 d,TSA、PTX、TSA+PTX和对照组Ark2细胞凋亡率膜联蛋白V染色法检测分别为4.25%±0.25%、12.12%±0.62%、16.56%±0.74%和46.78%±2.68%,Hoechst染色法检测分别为3.39%±0.12%、6.00%±0.25%、10.05%±0.53%和22.30%±1.25%,TSA+PIX组明显高于其他各组(均P＜0.05).药物作用后24 h,Ark2和KLE细胞的TSA+PTX组caspase-9、PARP的裂解明显增加;Ark7.和AN3细胞的TSA+PTX组MMP消失率(16.80%±0.92%、11.28%±0.78%)明显高于TSA组(4.96%±0.47%、6.46%±0.62%)和PTX组(5.34%±0.45%、5.61%±0.56%)(均P＜0.05);Ark2和KLE细胞的TSA+FIX组乙酰化微管蛋白表达明显增加.结论 TSA和PTX可通过促进子宫内膜癌细胞中乙酰化微管蛋白表达和增强微管稳定性而产生协同抗肿瘤作用.
目的 探討麯古抑菌素A(TSA)和紫杉醇(PTX)體外對人子宮內膜癌細胞凋亡和微管穩定性的影響及其機製.方法 將漿液性乳頭狀子宮內膜癌Ark2細胞、低分化子宮內膜樣腺癌KLE和AN3細胞各分為TSA、PTX單獨作用組(TSA組、PTX組)和聯閤作用組(TSA+PTX組),併設空白對照組,以錐蟲藍拒染法觀察藥物對細胞增殖的影響,膜聯蛋白V、Hoechst染色法檢測細胞凋亡率,流式細胞儀測定線粒體膜電位(MMP),蛋白質印跡法檢測半胱天鼕酶9(caspase-9)、多聚ADP覈糖聚閤酶(PARP)和乙酰化微管蛋白的錶達.結果 TSA、PTX對Ark2、KLE和AN3細胞均有抑製作用,二藥聯用後抑製作用更彊.藥物作用後4 d,TSA、PTX、TSA+PTX和對照組Ark2細胞凋亡率膜聯蛋白V染色法檢測分彆為4.25%±0.25%、12.12%±0.62%、16.56%±0.74%和46.78%±2.68%,Hoechst染色法檢測分彆為3.39%±0.12%、6.00%±0.25%、10.05%±0.53%和22.30%±1.25%,TSA+PIX組明顯高于其他各組(均P＜0.05).藥物作用後24 h,Ark2和KLE細胞的TSA+PTX組caspase-9、PARP的裂解明顯增加;Ark7.和AN3細胞的TSA+PTX組MMP消失率(16.80%±0.92%、11.28%±0.78%)明顯高于TSA組(4.96%±0.47%、6.46%±0.62%)和PTX組(5.34%±0.45%、5.61%±0.56%)(均P＜0.05);Ark2和KLE細胞的TSA+FIX組乙酰化微管蛋白錶達明顯增加.結論 TSA和PTX可通過促進子宮內膜癌細胞中乙酰化微管蛋白錶達和增彊微管穩定性而產生協同抗腫瘤作用.
목적 탐토곡고억균소A(TSA)화자삼순(PTX)체외대인자궁내막암세포조망화미관은정성적영향급기궤제.방법 장장액성유두상자궁내막암Ark2세포、저분화자궁내막양선암KLE화AN3세포각분위TSA、PTX단독작용조(TSA조、PTX조)화연합작용조(TSA+PTX조),병설공백대조조,이추충람거염법관찰약물대세포증식적영향,막련단백V、Hoechst염색법검측세포조망솔,류식세포의측정선립체막전위(MMP),단백질인적법검측반광천동매9(caspase-9)、다취ADP핵당취합매(PARP)화을선화미관단백적표체.결과 TSA、PTX대Ark2、KLE화AN3세포균유억제작용,이약련용후억제작용경강.약물작용후4 d,TSA、PTX、TSA+PTX화대조조Ark2세포조망솔막련단백V염색법검측분별위4.25%±0.25%、12.12%±0.62%、16.56%±0.74%화46.78%±2.68%,Hoechst염색법검측분별위3.39%±0.12%、6.00%±0.25%、10.05%±0.53%화22.30%±1.25%,TSA+PIX조명현고우기타각조(균P＜0.05).약물작용후24 h,Ark2화KLE세포적TSA+PTX조caspase-9、PARP적렬해명현증가;Ark7.화AN3세포적TSA+PTX조MMP소실솔(16.80%±0.92%、11.28%±0.78%)명현고우TSA조(4.96%±0.47%、6.46%±0.62%)화PTX조(5.34%±0.45%、5.61%±0.56%)(균P＜0.05);Ark2화KLE세포적TSA+FIX조을선화미관단백표체명현증가.결론 TSA화PTX가통과촉진자궁내막암세포중을선화미관단백표체화증강미관은정성이산생협동항종류작용.
Objective To investigate the effects of trichostatin A (TSA) and paclitaxel (PTX) on the apoptosis and microtubulin stabilization in human endometrial carcinoma cells and its mechanism. Methods Human endometrial carcinoma cells of the line Ark7., KLE and AN3 were cultured in the presence OfTSA (TSA group), or PTX(PTX group) , or TSA plus PTX (TSA +PTX group) respectively. The growth curve was obtained by trypan-blue exclusion assay. Apoptosis was observed by annexin V and Hoechst staining. Perturbation of mitochondrial membrane potential (MMP) was detected by flow cytometry. Western blotting was used to detect the protein expression of caspaae-9, poly ADP-riboae polymerase (PARP), and acetylated microtubulin. Results The growth of the Ark2, KLE, and AN3 cells of the TSA, PTX, and TSA + PTX group, especially in the latter group, was inhibited. The Ark2 cell apoptotic rates 4 days later of the TSA, PTX, TSA + FTX, and control group were 4.25%±0.25%, 12.12% ±0.62%, 16.56%±0.74%, and 46.78%±2.68% respectively by annexin V staining, and 3.39%±0.12%, 6.00%±0.25%, 10.05%±0.53%, and 22.30%±1.25% respectively by Hoechst staining. The apeptotie rates of the TSA + PTX group by both staining methods were both significantly higher than those of the other groups (all P＜0.05). Lysis of caspase-9 and PARP in the Ark2 and KLE cells increased greatly 24 hours after the TSA and PTX treatment. The disappearance rate of MMP in the Ark2 and AN3 cells of the TSA +PTX groups were 16.80%±0.92% and 11.28%±0.78% respectively, significandy higher than that of the PTX group (5.34%±0.45% and 5.61%±0.56% respectively) and TSA group (4.96%± 0.47% and 6.46%±0.62% respectively, all P＜0.05). The expression of acetylated mieretubulin was increased in the Ark2 and KLE cells of the TSA + PTX groups. Condusions Synergy of TSA and PTX inhibits the cell growth and induces apoptosis. The acetylation of non-histone protein induced by histone deacetylase inhibitor is one of the possible mechanisms of its anti-cancer effects.
