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??OBJECTIVE To screen the best combination of extractum of Robinia-living trametes and chemotherapy and investigate the action mechanism of Robinia-living trametes against the apoptosis of human gastric cancer cell MGC803. METHODS MGC803 Cells were treated with different concentrations of Robinia-living trametes and chemotherapy drugs (5-Fu and paclitaxel) in vitro. The inhibitory rate of cells was measured by MTT assay. Morphological changes were observed with inverted microscope. The apoptosis rate of MGC803 cells which were treated with combination of Robinia-living trametes(0.2 mgmL^-1) and 5-Fu (2.5 ??gmL^-1) was detected by FCM. The protein expression of P53 and p-Akt in MGC803 cells which were treated with combination of Robinia-living trametes (0.2 mgmL^-1) and 5-Fu (2.5 ??gmL^-1) was detected by Western blot. RESULTS The viability of MGC803 cells was reduced by Robinia-living trametes and chemotherapy drugs (5-Fu and paclitaxel) in a concentration- and time-dependent manner(P<0.01). Under reverse microscopy, cell body shrinking, nuclear pyrosis, and nuclear fragmentation were observed. The higher concentration, the longer treatment time, the more cells died. Compared with monotherapy, the combination of Robinia-living trametes and chemotherapy could reduce the survival rate of MGC803 cells. The protein expressions of P53 in MGC803 cells treated with combination of drugs was up-regulated, while that of P-Akt was down-regulated. CONCLUSION The apoptosis of MGC803 cells in vitro may be induced by the inhibitory effect of the combination of Robinia-living trametes and 5-Fu on PI3K/Akt signaling pathway. Combination therapy of Robinia-living trametes and 5-Fu is potentially more effective in inhibition of tumor cells than monotherapy of Robinia-living trametes.     

健脾消癥方对人胃癌细胞MGC803凋亡和细胞周期的影响

目的:观察健脾消癥方对胃癌细胞株MGC803增殖、凋亡和细胞周期的影响。方法:(1)采用MTT法观察不同浓度的健脾消癥方对胃癌细胞株MGC803增殖的抑制作用;(2)使用Annexin V/PI荧光双染法检测健脾消癥方诱导肿瘤细胞进入凋亡的比例;(3)应用流式细胞仪检测健脾消癥方对MGC803细胞周期的影响。结果:(1)健脾消癥方对MGC803细胞有较强的抑制增殖的作用,并呈一定的浓度依赖性。(2)健脾消癥方高、中、低浓度组对胃癌细胞株MGC803作用24h后,早期凋亡率分别为49.83%、55.29%、16.73%。健脾消癥方高、中、低浓度组对胃癌细胞株MGC803作用48h后,早期凋亡率分别为84.52%、55.58%、51.76%。(3)与阴性对照组相比,健脾消癥方各浓度组对胃癌细胞株MGC803作用48h后,可将细胞阻滞于G2-M期,健脾消癥方高浓度组在G0-G1期前出现较明显的亚二倍体峰。结论:(1)健脾消癥方可明显抑制胃癌细胞株MGC803的增殖作用。(2)健脾消癥方对人胃癌细胞株MGC803有明显的诱导凋亡及阻滞细胞周期在G2-M期的作用。     

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??OBJECTIVE To investigate the effects of combination treatment with L-carnitine and 5-fluorouracil on the proliferation and cell apoptosis of gastric cancer MGC803 cells. METHODS MGC803 cells were divided into control group, 5-fluorouracil group and the combination of L-carnitine and 5-fluorouracil group (L-carnitine+/??5-fluorouracil group) in vitro. The inhibitory rate of cells was measured by MTT assay. The apoptosis rate and cell cycle of cells were detected by FLOW. Western blot was used to analyzed the expression of Bcl-2, Bax, adenine nucleotide translocator1(ANT1) and cleaved-PARP. RESULTS Compared with 5-fluorouracil group, the inhibition rate of MGC803 cells was increased when cells were treated with the combination of L-carnitine and 5-fluorouracil. The apoptosis rate of cells was raised and the cells were blocked at S phase. In addition, the combination group can decrease the expression of Bcl-2 and increase the expression of Bax, ANT1 and cleaved-PARP. At the same time, the apoptosis rate of cells and the cell cycle were different with the different dosage regimen when treated with the combination. Compared with the L-carnitine+5-fluorouracil group, the apoptosis rate of cells was increased to (24.17??3.12)% from (19.60??1.06)% (P<0.05). The G₀/G₁ phase proportion of cells was decreased to (62.62??1.04)% from (72.95??0.91)%,and the S phase proportion of cells was increased to (37.35??1.03)% from (27.05??0.91)% (P<0.001). CONCLUSION Treatment with L-carnitine and 5-fluorouracil could enhance the inhibitory effect of 5-fluorouracil on MGC803 cells. The possible mechanism of action is achieved by regulating the expression of Bcl-2 protein family and influencing the cell cycle.     

紫杉醇与白藜芦醇联合应用对人胃癌MGC803细胞作用的研究

目的:研究紫杉醇(PA)联合白藜芦醇(RES)对人胃癌MGC803细胞株的影响.方法:采用MTT法检测两种药物单用和联合应用对MGC 803细胞的作用,光镜观察细胞结构变化,流式细胞术检测细胞周期分布.结果:PA可抑制MGC803细胞生长,呈剂量及时间依赖性(P<0.05);PA促进人胚肺成纤维细胞(HPF)生长,呈剂量依赖性(P<0.05);PA与RES联合应用对MGC803生长抑制有协同作用(P<0.05),光镜下细胞质内产生异常物质;PA对G0/G1期有阻滞作用,RES对S期有明显阻滞作用.结论:PA对胃癌MGC803细胞有抑制作用.PA与RES对胃癌MGC 803细胞抑制具有协同作用,与单用RES相比,PA与RES联合应用可减少RES的用药剂量.     

芪竹方中单体及配伍对人胃癌细胞MGC-803抑制作用研究

目的:观察芪竹方单体成分薯蓣皂苷、莪术醇及两者配伍对人胃癌细胞MGC-803的增殖抑制作用,初步探讨芪竹方复方在抗肿瘤治疗中的有效成分及作用机制。方法:选用5个剂量的薯蓣皂苷、莪术醇单药组,及2个剂量的两药配伍组在24h、48h 2个不同时间点作用人胃癌细胞MGC-803,采用MTT法检测药物对MGC-803的增殖抑制率。结果:一定浓度范围内,薯蓣皂苷及莪术醇均对MGC-803细胞的增殖有剂量、时间依赖性抑制作用,且两药配伍使用较单药能产生更好的抑制作用。结论:薯蓣皂苷及莪术醇可能为芪竹方抑制人胃癌细胞MGC-803增殖的有效成分。     

一株连翘内生真菌代谢粗提物对胃癌细胞株MGC-803增殖的影响

目的：评价一株连翘内生真菌代谢粗提物体外对胃癌细胞株MGC-803增殖的影响,为寻找临床胃癌治疗新药提供思路。方法：采用常规分离手段分离得到连翘内生真菌26株,应用MTT法测定其发酵代谢产物粗提物对MGC-803增殖的影响。结果：MGC-803经内生真菌FS12的发酵代谢产物粗提物FE-12作用后细胞克隆受到明显的抑制,且在时间和剂量上存在一定的依赖性。结论：连翘内生真菌FS12的发酵粗提物FE-12可抑制胃癌细胞株MGC-803增殖,为寻找临床胃癌治疗新药提供了一条新的思路。     

川陈皮素对乳腺癌细胞的化疗增敏作用

目的 探讨川陈皮素对乳腺癌细胞MDA-MB-231的体外化疗增敏作用.方法 以MDA-MB-231细胞为研究对象,MTT法测定细胞增殖抑制率,DNA Ladder、核小体双抗体ELISA和流式细胞仪测定MDA-MB-231细胞凋亡情况,凝胶阻滞实验(EMSA)和ELISA检测川陈皮素对核转录因子(NF-κB)的转录激活的影响.结果 20 μmol/L川陈皮素分别与1 nmol/L紫杉醇或50 ng/mL阿霉素联合使用肿瘤细胞增殖抑制率为75.1%和82.6%,单独使用1 nmol/L紫杉醇或50 ng/mL阿霉素肿瘤细胞增殖抑制率为40%和45%;核小体双抗体夹心酶免疫法和DNA凝胶电泳实验表明川陈皮素与紫杉醇或阿霉素联合用药可明显促进MDA-MB-231细胞的凋亡,EMSA和ELISA实验表明川陈皮素与紫杉醇或阿霉素联合用药可明显抑制NF-κB的转录活性.结论 川陈皮素可以促进临床化疗药物对乳腺癌细胞增殖的抑制作用和诱导凋亡作用,其机制可能是通过抑制NF-κB转录作用.     

姜黄素抑制人胃癌BGC-823细胞生长并诱导其凋亡的机制研究

目的:探讨姜黄素抑制人胃癌BGC-823细胞生长并促人胃癌BGC-823细胞凋亡的生物学作用及其调控机制。方法:常规体外培养对数生长期胃癌BGC-823细胞,细胞分为对照组,低、中、高姜黄素处理组四组,姜黄素浓度分别为0mg/L5,mg/L1,0mg/L,20mg/L。姜黄素处理24h后,采用甲基噻唑(MTT)比色法及流式细胞仪测定细胞增殖水平及细胞凋亡率;采用免疫组化法测定细胞内Bax、Bcl-2蛋白表达;采用PCR检测Caspase-3的mRNA表达水平。结果:MTT检测显示姜黄素能抑制人胃癌BGC-823细胞増殖,呈现浓度依赖性;流式细胞仪显示姜黄素能有效诱导细胞的凋亡,呈现浓度依赖性,其中20mg/L姜黄素处理24h后细胞凋亡率为48.3%;免疫组化试验表明姜黄素处理使人胃癌BGC-823细胞中Bax表达水平上调,同时Bcl-2蛋白表达水平下调;且细胞中Caspase-3的mRNA表达水平受姜黄素诱导而增高。结论:姜黄素对人胃癌BGC-823细胞的增殖具有明显抑制作用,呈浓度依赖性促进细胞凋亡,这种生物学效应可能与激活Bax蛋白表达、抑制Bcl-2蛋白表达而活化Caspase-3的信号通路有关。该研究为深入探讨姜黄素诱导人胃癌BGC-823细胞凋亡的机制提供了重要依据。     

藏药柳茶水提物对胃癌耐药细胞BGC823/5-FU增殖及药物敏感性的影响

研究藏药柳茶水提物(Liu Tea extracts,LTE)对人胃癌氟尿嘧啶耐药细胞株BGC823/5-FU的增殖和药物敏感性的影响。将LTE作用于人胃癌耐药细胞BGC823/5-FU,采用MTT检测其对细胞增殖、化疗药物敏感性的影响以及与5-氟尿嘧啶(5-FU)的联用效应,应用CompuSyn软件计算联合用药指数(combination index,CI);流式细胞术(FCM)检测LTE对BGC823/5-FU细胞凋亡的作用;Western blot检测不同质量浓度(100,200,400 mg·L-1)LTE对5-FU作用下胃癌耐药细胞中P-gp,Bcl-2,Bax及Caspase-3(17KD)蛋白表达的影响。结果显示,LTE能够有效的抑制胃癌耐药细胞株BGC823/5-FU的生长,抑制率与药物浓度及作用时间呈正相关;LTE能够促进细胞发生凋亡,800 mg·L-1的LTE作用细胞24 h后凋亡率可高达46.2%(P0.01)。LTE与5-FU联用后CI1,说明两药具有较好的协同抑制耐药细胞增殖的作用。非细胞毒性剂量LTE可显著降低化疗药物5-FU,CDDP,PTX,ADM对BGC823/5-FU细胞的半数抑制浓度(IC50,P0.05),一定程度上逆转耐药细胞对化疗药物的耐药性,逆转倍数分别为2.35,1.68,1.96,0.52。蛋白检测结果显示,随着柳茶水提物浓度的增加,耐药细胞内Bcl-2表达逐渐下降,Bax和Caspase-3蛋白表达显著升高,差异具有统计学意义(P0.01)。但耐药蛋白P-gp的表达在实验组和对照组细胞中无显著变化。LTE能有效抑制胃癌耐药细胞株BGC823/5-FU的生长,并在一定程度上逆转其化疗耐药。抑制抗凋亡蛋白的表达、激活促凋亡蛋白、诱导耐药细胞发生凋亡可能是其作用的主要机制。