中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
12期
2897-2899
,共3页
茆勇%马凤锦%刘合勇%黄朝晖%游庆军%华东
茆勇%馬鳳錦%劉閤勇%黃朝暉%遊慶軍%華東
묘용%마봉금%류합용%황조휘%유경군%화동
顺铂%唑来膦酸%A549细胞%DNA损伤检查点蛋白调节因子1%DNA切除修复交叉互补基因1%受体相关蛋白80基因
順鉑%唑來膦痠%A549細胞%DNA損傷檢查點蛋白調節因子1%DNA切除脩複交扠互補基因1%受體相關蛋白80基因
순박%서래련산%A549세포%DNA손상검사점단백조절인자1%DNA절제수복교차호보기인1%수체상관단백80기인
Cisplatin%Zoledronic acid%A549 cell%Mediator of DNA damage checkpoint protein 1%Excision repair cross complementation group 1%Receptor-associated protein 80
目的 比较唑来膦酸与顺铂异序联合对肺腺癌A549细胞的抑制作用.方法 按照唑来膦酸和顺铂药物处理的不同顺序将肺腺癌A549细胞分为4组(唑来膦酸和顺铂的药物浓度分别为1.560 0 mmol/L和0.312 5mg/L):(1)A组(对照组):肺腺癌A549细胞常规培养,不作药物处理;(2)B组(同时用药组):同时加入唑来膦酸和顺铂孵育48 h;(3)C组(顺铂+唑来膦酸组):顺铂孵育24 h后,加入唑来膦酸孵育24 h;(4)D组(唑来膦酸+顺铂组):唑来膦酸孵育24 h后,加入顺铂孵育24 h.采用噻唑蓝(MTT)法检测各组细胞体外增殖水平;膜联蛋白V(Annexin V)/碘化丙锭(PI)双染色法检测各组细胞凋亡;实时荧光定量聚合酶链反应(FQ-PCR)法检测各组细胞DNA损伤检查点蛋白调节因子1(MDC1)、DNA切除修复交叉互补基因1(ERCC1)和受体相关蛋白80(RAP80)基因mRNA的表达.结果 A、B、C、D组细胞生长抑制率分别为0、(39.16±4.94)％、(46.94±3.45)％、(38.43±4.84)％,B、C、D组均显著高于A组(P＜0.05),B、D组均显著低于C组(P＜0.05);各组细胞凋亡率分别为(0.53±0.15)％、(32.3±0.50)％、(36.53±0.31)％、(29.33±2.48)％,B、C、D组均显著高于A组(P＜0.05),B、D组均显著低于C组(均P ＜0.05);B、C、D组细胞中MDC1和RAP80基因mRNA的表达(0.374±0.022和1.127±0.018、0.320±0.023和1.036±0.0279.416±0.016和1.181±0.041)均显著低于A组(0.677±0.028和1.548±0.038-P＜0.05),且B、D组均显著高于C组(P＜0.05);各组细胞ERCCl基因mRNA表达差异无统计学意义(P＞0.05).结论 顺铂联合唑来膦酸可抑制肺癌A549细胞的增殖,诱导其凋亡,并有顺序依赖性,以顺铂序以唑来膦酸效果更显著;其作用可能与下调MDC1和RAP80基因mRNA表达有关.
目的 比較唑來膦痠與順鉑異序聯閤對肺腺癌A549細胞的抑製作用.方法 按照唑來膦痠和順鉑藥物處理的不同順序將肺腺癌A549細胞分為4組(唑來膦痠和順鉑的藥物濃度分彆為1.560 0 mmol/L和0.312 5mg/L):(1)A組(對照組):肺腺癌A549細胞常規培養,不作藥物處理;(2)B組(同時用藥組):同時加入唑來膦痠和順鉑孵育48 h;(3)C組(順鉑+唑來膦痠組):順鉑孵育24 h後,加入唑來膦痠孵育24 h;(4)D組(唑來膦痠+順鉑組):唑來膦痠孵育24 h後,加入順鉑孵育24 h.採用噻唑藍(MTT)法檢測各組細胞體外增殖水平;膜聯蛋白V(Annexin V)/碘化丙錠(PI)雙染色法檢測各組細胞凋亡;實時熒光定量聚閤酶鏈反應(FQ-PCR)法檢測各組細胞DNA損傷檢查點蛋白調節因子1(MDC1)、DNA切除脩複交扠互補基因1(ERCC1)和受體相關蛋白80(RAP80)基因mRNA的錶達.結果 A、B、C、D組細胞生長抑製率分彆為0、(39.16±4.94)％、(46.94±3.45)％、(38.43±4.84)％,B、C、D組均顯著高于A組(P＜0.05),B、D組均顯著低于C組(P＜0.05);各組細胞凋亡率分彆為(0.53±0.15)％、(32.3±0.50)％、(36.53±0.31)％、(29.33±2.48)％,B、C、D組均顯著高于A組(P＜0.05),B、D組均顯著低于C組(均P ＜0.05);B、C、D組細胞中MDC1和RAP80基因mRNA的錶達(0.374±0.022和1.127±0.018、0.320±0.023和1.036±0.0279.416±0.016和1.181±0.041)均顯著低于A組(0.677±0.028和1.548±0.038-P＜0.05),且B、D組均顯著高于C組(P＜0.05);各組細胞ERCCl基因mRNA錶達差異無統計學意義(P＞0.05).結論 順鉑聯閤唑來膦痠可抑製肺癌A549細胞的增殖,誘導其凋亡,併有順序依賴性,以順鉑序以唑來膦痠效果更顯著;其作用可能與下調MDC1和RAP80基因mRNA錶達有關.
목적 비교서래련산여순박이서연합대폐선암A549세포적억제작용.방법 안조서래련산화순박약물처리적불동순서장폐선암A549세포분위4조(서래련산화순박적약물농도분별위1.560 0 mmol/L화0.312 5mg/L):(1)A조(대조조):폐선암A549세포상규배양,불작약물처리;(2)B조(동시용약조):동시가입서래련산화순박부육48 h;(3)C조(순박+서래련산조):순박부육24 h후,가입서래련산부육24 h;(4)D조(서래련산+순박조):서래련산부육24 h후,가입순박부육24 h.채용새서람(MTT)법검측각조세포체외증식수평;막련단백V(Annexin V)/전화병정(PI)쌍염색법검측각조세포조망;실시형광정량취합매련반응(FQ-PCR)법검측각조세포DNA손상검사점단백조절인자1(MDC1)、DNA절제수복교차호보기인1(ERCC1)화수체상관단백80(RAP80)기인mRNA적표체.결과 A、B、C、D조세포생장억제솔분별위0、(39.16±4.94)％、(46.94±3.45)％、(38.43±4.84)％,B、C、D조균현저고우A조(P＜0.05),B、D조균현저저우C조(P＜0.05);각조세포조망솔분별위(0.53±0.15)％、(32.3±0.50)％、(36.53±0.31)％、(29.33±2.48)％,B、C、D조균현저고우A조(P＜0.05),B、D조균현저저우C조(균P ＜0.05);B、C、D조세포중MDC1화RAP80기인mRNA적표체(0.374±0.022화1.127±0.018、0.320±0.023화1.036±0.0279.416±0.016화1.181±0.041)균현저저우A조(0.677±0.028화1.548±0.038-P＜0.05),차B、D조균현저고우C조(P＜0.05);각조세포ERCCl기인mRNA표체차이무통계학의의(P＞0.05).결론 순박연합서래련산가억제폐암A549세포적증식,유도기조망,병유순서의뢰성,이순박서이서래련산효과경현저;기작용가능여하조MDC1화RAP80기인mRNA표체유관.
Objective To compare lung cancer A549 cell inhibition treated by zoledronic acid different schedule combined with cisplatin.Methods According to different schedule of drug treatment,lung cancer A549 cells were divided into 4 groups (drug concentrations of zoledronic acid and cisplatin were 1.560 0,0.312 5 mg/L):(1) A group (control group):A549 cells were cultured,no drug treatment; (2) B group (simultaneously treated group):zoledronic acid and cisplatin were added simultaneously; (3) C group (cisplatin + zoledronic acid group):cells were incubated with cisplatin for 24 h followed by with zoledronic acid for 24 h; (4) D group (zoledronic acid + cisplatin group):cells were incubated with zoledronic acid for 24 h followed by with cisplatin.Methyl thiazol tetrazolium (MTT) assay and annexin V/ propidium iodide (PI) double staining were used to examine each groups A549 cells proliferation and apoptosis respectively.Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was applied to detected mRNA expression of mediator of DNA damage checkpoint protein 1 (MDC1),excision repair cross complementation group 1 (ERCC1) and receptor-associated protein 80 (RAP80) in A549 cells.Results Inhibition rates of A,B,C,D groups A549 cells were 0％,(39.16±4.94)％,(46.94 ±3.45) ％,(38.43 ± 4.84) ％,respectively.Rates of B,C,D groups were significantly higher than that of A group (all P ＜ 0.05),Rates of B,D group were significantly lower than that of C group (all P ＜ 0.05) ; Apoptosis rates of each group were (0.53 ±0.15)％,(32.3 ±0.50)％,(36.53 ±0.31)％,(29.33 ± 2.48) ％,respectively.Rates of B,C,D groups were significantly higher than that of A group (all P ＜ 0.05).Rates ofB,D group were significantly lower than that of C group (all P＜0.05); MDC1 and RAP80 gene expressions of B,C,D groups (0.374 ± 0.022 and 1.127 ± 0.018,0.320 ± 0.023 and 1.036 ±0.027,0.416 ±0.016 and 1.181 ±0.041,respectively) were significantly lower than that of A group (0.677 ±0.028 and 1.548 ±0.038) (all P ＜0.05),Expressions of B,D group were significantly higher than that of C group (all P ＜ 0.05) ; Expressions of ERCC1 mRNA had no significantly difference (all P ＞ 0.05).Conclusion Zoledronic acid combined with cisplatin can inhibit lung cancer A549 cells proliferation and induce cells apoptosis synergistically with schedule-dependent.The most remarkable effect was found in cells treated with cisplatin followed by with zoledronic acid.The downregulated expression of MDC 1 and RAP80 may be involved in the mechanism.
