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
1416-1418
,共3页
扈及雷%王修杰%周培志%马卫朝%姜曙
扈及雷%王脩傑%週培誌%馬衛朝%薑曙
호급뢰%왕수걸%주배지%마위조%강서
共轭三烯酸%胶质瘤%增殖
共軛三烯痠%膠質瘤%增殖
공액삼희산%효질류%증식
Conjugated trienoic fatty acids%Glioma%Proliferation
目的 探讨共轭三烯酸(TCLA)对人胶质瘤细胞的增殖抑制作用及作用机制。方法 用噻唑蓝(MTT)比色法、克隆形成试验、5-溴脱氧尿嘧啶核苷(BrdU)掺人试验研究TCLA对人脑胶质瘤细胞的增殖抑制作用;Hoechst 33342/PI双染法、流式细胞术检测细胞凋亡指数和周期分布;逆转录-聚合酶链反应(RT-PCR)检测凋亡相关基因腺苷二磷酸核糖转移酶1(ADPRTL1)、细胞色素P4501A1( CYP1A1),过氧化物酶增殖体激活受体-γ(PPAR-γ)的表达。结果 共轭三烯酸对人胶质细胞瘤(U251)有明显抑制作用,呈剂量-时间-效应关系(P＜0.05)。克隆形成下降,40 μmol/LTCLA可完全抑制克隆形成。BrdU标记从(91.6±3.6)％下降到(14.4±4.4)％(P＜0.05),Hoechst 33342/PI双染试验,凋亡细胞增加(P＜0.05)。流式细胞仪检测显示,细胞凋亡指数从(7.3±1.2)％升高到(34.2±2.4)％(P＜0.05),Go/G1期细胞比例增加,S期细胞比例减少(P＜0.05);RT-PCR结果显示,凋亡相关基因ADPRTL1、CYP1 A1、PPAR-γ mRNA表达增强(P＜0.05)。结论 TCIA对人脑胶质瘤细胞具有增殖抑制和凋亡诱导作用,其作用机制可能与抑制肿瘤细胞DNA合成、细胞周期阻滞、上调凋亡相关基因(ADPRTL1、CYP1A1、PPAR-γ)表达有关。
目的 探討共軛三烯痠(TCLA)對人膠質瘤細胞的增殖抑製作用及作用機製。方法 用噻唑藍(MTT)比色法、剋隆形成試驗、5-溴脫氧尿嘧啶覈苷(BrdU)摻人試驗研究TCLA對人腦膠質瘤細胞的增殖抑製作用;Hoechst 33342/PI雙染法、流式細胞術檢測細胞凋亡指數和週期分佈;逆轉錄-聚閤酶鏈反應(RT-PCR)檢測凋亡相關基因腺苷二燐痠覈糖轉移酶1(ADPRTL1)、細胞色素P4501A1( CYP1A1),過氧化物酶增殖體激活受體-γ(PPAR-γ)的錶達。結果 共軛三烯痠對人膠質細胞瘤(U251)有明顯抑製作用,呈劑量-時間-效應關繫(P＜0.05)。剋隆形成下降,40 μmol/LTCLA可完全抑製剋隆形成。BrdU標記從(91.6±3.6)％下降到(14.4±4.4)％(P＜0.05),Hoechst 33342/PI雙染試驗,凋亡細胞增加(P＜0.05)。流式細胞儀檢測顯示,細胞凋亡指數從(7.3±1.2)％升高到(34.2±2.4)％(P＜0.05),Go/G1期細胞比例增加,S期細胞比例減少(P＜0.05);RT-PCR結果顯示,凋亡相關基因ADPRTL1、CYP1 A1、PPAR-γ mRNA錶達增彊(P＜0.05)。結論 TCIA對人腦膠質瘤細胞具有增殖抑製和凋亡誘導作用,其作用機製可能與抑製腫瘤細胞DNA閤成、細胞週期阻滯、上調凋亡相關基因(ADPRTL1、CYP1A1、PPAR-γ)錶達有關。
목적 탐토공액삼희산(TCLA)대인효질류세포적증식억제작용급작용궤제。방법 용새서람(MTT)비색법、극륭형성시험、5-추탈양뇨밀정핵감(BrdU)참인시험연구TCLA대인뇌효질류세포적증식억제작용;Hoechst 33342/PI쌍염법、류식세포술검측세포조망지수화주기분포;역전록-취합매련반응(RT-PCR)검측조망상관기인선감이린산핵당전이매1(ADPRTL1)、세포색소P4501A1( CYP1A1),과양화물매증식체격활수체-γ(PPAR-γ)적표체。결과 공액삼희산대인효질세포류(U251)유명현억제작용,정제량-시간-효응관계(P＜0.05)。극륭형성하강,40 μmol/LTCLA가완전억제극륭형성。BrdU표기종(91.6±3.6)％하강도(14.4±4.4)％(P＜0.05),Hoechst 33342/PI쌍염시험,조망세포증가(P＜0.05)。류식세포의검측현시,세포조망지수종(7.3±1.2)％승고도(34.2±2.4)％(P＜0.05),Go/G1기세포비례증가,S기세포비례감소(P＜0.05);RT-PCR결과현시,조망상관기인ADPRTL1、CYP1 A1、PPAR-γ mRNA표체증강(P＜0.05)。결론 TCIA대인뇌효질류세포구유증식억제화조망유도작용,기작용궤제가능여억제종류세포DNA합성、세포주기조체、상조조망상관기인(ADPRTL1、CYP1A1、PPAR-γ)표체유관。
Objective To investigate the effect of conjugated trienoic fatty acids (TCLA) on proliferation of human glioblastoma U251 cells and the action mechanism. Methods Glioblastoma cells were tested in vitro by methyl thiazol tetrazolium ( MTT), colony formation inhibition, BrdU incorporation after treatment with TCLA. The apoptotic cells were assayed through Hoechst 33342/propidium iodide staining and cell cycle analysis by flow cytometry (FCM), and the expression of ADPRTL1, CYP1A1 and PPAR-γgenes were detected by using reverse transcription-polymerase chain reaction (RT-PCR). Results After TCLA treatment, the proliferation of U251 cells was inhibited in a dose-and time-dependent manner (P ＜0. 05), colony formation decreased significantly (P ＜ 0. 05 ) and BrdU labeling index of cancer cells decreased from (91.6 ± 3.6) ％ to ( 14. 4 ± 4. 4) ％ ( P ＜ 0. 05 ) ; apoptotic cells increased; FCM revealed that the apoptotic indices were increased from (7. 3 ± 1.2) ％ to (34. 2 ± 2. 4) ％ ( P ＜ 0. 05 ), the cells in G0/G1 phase were increased, and those in S phase decreased (P ＜ 0. 05). RT-RCR showed that TCLA significantly increased the mRNA expression levels of ADPRTL1, CYP1A1, and PPAR-γ. Conclusion The findings in this experimental study suggested that TCLA had potent cytotoxicity and apoptosis induction effect on human glioblastoma U251 cells probably by inhibiting DNA synthesis, blocking cell cycle and upregulating the expression of apoptosis related genes ADPRTL1, CYP1A1, PPAR-γ.
