中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
9期
1526-1529
,共4页
粉防己碱%阿霉素%膀胱癌%抗药性
粉防己堿%阿黴素%膀胱癌%抗藥性
분방기감%아매소%방광암%항약성
Tetrandrine%Adriamycin%Bladder carcinoma%Drug resisitance
目的 探讨粉防己碱(TET)对膀胱癌耐药株BIU-87/ADM多药耐药性(MDR)的逆转作用及其机制。方法 应用CCK法观察不同浓度TET( 1.0、1.5、2.0、2.5 mg/L)对BIU-87和BIU-87/ADM细胞生长的抑制作用,并测定1 mg/L TET逆转多药耐药的活性;Annexin-V与PI双染法流式细胞术分析1 mg/L TET对细胞凋亡的影响;免疫荧光法和逆转录-聚合酶链反应(RT-PCR)分别检测1 mg/L TET对P-gp蛋白和MDR1表达水平的影响;细胞免疫化学法和RT-PCR法检测1 mg/L TET对BIU-87/ADM细胞Caspase-3和Survivin表达水平的影响。结果 1.0 mg/L无毒剂量下的TET显著增加阿霉素(ADM)对耐药株BIU-87/ADM的细胞毒性作用(P＜0.01),逆转指数(RF)为5.33,而对敏感株BIU-87无明显作用(P＞0.05),RF为1.33;1.0 mg/L TET和1.5 mg/LADM共同作用48 h,能使BIU-87/ADM细胞凋亡率增加至(40.86±4.35)％,与单独应用ADM的(12.42 ±3.24)％比较差异有统计学意义(P＜0.01)。而BIU-87细胞凋亡率为(61.23 ±5.46)％,与单独应用ADM的(57.79±4.73)％比较差异无统计学意义(P＞0.05)。TET能明显抑制mdr1mRNA和P-gp蛋白在BIU-87/ADM的表达;与单用ADM比较,TET和ADM联用能使BIU-87/ADM细胞株中Caspase-3的表达水平明显升高(P＜0.01),Survivin的表达水平显著降低(P＜0.01)。结论 粉防己碱能逆转BIU-87/ADM细胞的多药耐药性,这一作用可能与粉防己碱抑制P-gp的表达和增强化疗药诱导细胞凋亡有关。
目的 探討粉防己堿(TET)對膀胱癌耐藥株BIU-87/ADM多藥耐藥性(MDR)的逆轉作用及其機製。方法 應用CCK法觀察不同濃度TET( 1.0、1.5、2.0、2.5 mg/L)對BIU-87和BIU-87/ADM細胞生長的抑製作用,併測定1 mg/L TET逆轉多藥耐藥的活性;Annexin-V與PI雙染法流式細胞術分析1 mg/L TET對細胞凋亡的影響;免疫熒光法和逆轉錄-聚閤酶鏈反應(RT-PCR)分彆檢測1 mg/L TET對P-gp蛋白和MDR1錶達水平的影響;細胞免疫化學法和RT-PCR法檢測1 mg/L TET對BIU-87/ADM細胞Caspase-3和Survivin錶達水平的影響。結果 1.0 mg/L無毒劑量下的TET顯著增加阿黴素(ADM)對耐藥株BIU-87/ADM的細胞毒性作用(P＜0.01),逆轉指數(RF)為5.33,而對敏感株BIU-87無明顯作用(P＞0.05),RF為1.33;1.0 mg/L TET和1.5 mg/LADM共同作用48 h,能使BIU-87/ADM細胞凋亡率增加至(40.86±4.35)％,與單獨應用ADM的(12.42 ±3.24)％比較差異有統計學意義(P＜0.01)。而BIU-87細胞凋亡率為(61.23 ±5.46)％,與單獨應用ADM的(57.79±4.73)％比較差異無統計學意義(P＞0.05)。TET能明顯抑製mdr1mRNA和P-gp蛋白在BIU-87/ADM的錶達;與單用ADM比較,TET和ADM聯用能使BIU-87/ADM細胞株中Caspase-3的錶達水平明顯升高(P＜0.01),Survivin的錶達水平顯著降低(P＜0.01)。結論 粉防己堿能逆轉BIU-87/ADM細胞的多藥耐藥性,這一作用可能與粉防己堿抑製P-gp的錶達和增彊化療藥誘導細胞凋亡有關。
목적 탐토분방기감(TET)대방광암내약주BIU-87/ADM다약내약성(MDR)적역전작용급기궤제。방법 응용CCK법관찰불동농도TET( 1.0、1.5、2.0、2.5 mg/L)대BIU-87화BIU-87/ADM세포생장적억제작용,병측정1 mg/L TET역전다약내약적활성;Annexin-V여PI쌍염법류식세포술분석1 mg/L TET대세포조망적영향;면역형광법화역전록-취합매련반응(RT-PCR)분별검측1 mg/L TET대P-gp단백화MDR1표체수평적영향;세포면역화학법화RT-PCR법검측1 mg/L TET대BIU-87/ADM세포Caspase-3화Survivin표체수평적영향。결과 1.0 mg/L무독제량하적TET현저증가아매소(ADM)대내약주BIU-87/ADM적세포독성작용(P＜0.01),역전지수(RF)위5.33,이대민감주BIU-87무명현작용(P＞0.05),RF위1.33;1.0 mg/L TET화1.5 mg/LADM공동작용48 h,능사BIU-87/ADM세포조망솔증가지(40.86±4.35)％,여단독응용ADM적(12.42 ±3.24)％비교차이유통계학의의(P＜0.01)。이BIU-87세포조망솔위(61.23 ±5.46)％,여단독응용ADM적(57.79±4.73)％비교차이무통계학의의(P＞0.05)。TET능명현억제mdr1mRNA화P-gp단백재BIU-87/ADM적표체;여단용ADM비교,TET화ADM련용능사BIU-87/ADM세포주중Caspase-3적표체수평명현승고(P＜0.01),Survivin적표체수평현저강저(P＜0.01)。결론 분방기감능역전BIU-87/ADM세포적다약내약성,저일작용가능여분방기감억제P-gp적표체화증강화료약유도세포조망유관。
Objective To Investigate the reversal effect and the mechanism of tetrandrine (TET) on human bladder carcinoma cell line BIU-87/ADM. Methods The effects of TET ( 1.0, 1.5, 2. 0, 2. 5mg/L) on the growth of BIU-87 and BIU-87/ADM cells were measured by CCK assay. The potential of 1 mg/L TET to reverse multidrug resisitance (MDR) was also determined by CCK assay. The apoptosis of cell lines induced by 1 mg/L TET was evaluated by Annexin-V and PI followed by flow cytometry (FCM).The effects of 1 mg/L TET on the expression of P-gp and MDR1 were separately measured by immunofluorescence and reverse transcription-polymerase chain reaction (RT-PCR), and the expression of Caspase-3 and Survivin was analyzed by immunocytochemical technique and RT-PCR. Results TET at the concentration of 1.0 mg/L which had no obvious cytotoxicity to the BIU-87 and BIU-87/ADM cells increased the cytotoxicity of adriamycin (ADM) in BIU-87/ADM cells greatly, with a reversal fold of 5. 33, but had no significant effect on BIU-87 cells, with a reversal fold of 1.33. The ADM-induced apoptosis rate in BIU-87 and BIU-87/ADM cells was (57. 79 ± 4. 73 ) ％ and ( 12.42 ± 3.24 ) ％ respectively. After treatment of BIU-87/ADM cells with 1.0 mg/L TET, the apoptosis rate was increased to (40. 86 ±4. 35)％, but it made no difference in BIU-87 cells. The expression of mdr1 and P-gp was suppressed by TET in BIU-87/ADM cells. Compared to the ADM group, Caspase-3 was more highly expressed in BIU-87/ADM cells of ADM +TET group. However, Survivin had the opposite result. Conclusion TET can reverse the MDR of the human bladder carcinoma BIU-87/ADM cell line, which may be associated with the decreased expression of P-gp and increased apoptosis.
