奥曲肽联合wortmannin对人胃癌细胞BGC-823生长抑制作用的研究  

Effects of combination of octreotide and wortmannin on proliferation of gastric carcinoma cell BGC-823

作　　者：张利[1] 于红刚[1] 于皆平[1] 罗和生[1] 

机构地区：[1]武汉大学人民医院消化科,武汉430060

出　　处：《中国实用内科杂志》2008年第3期180-183,共4页Chinese Journal of Practical Internal Medicine

基　　金：国家自然科学基金(30300154)

摘　　要：目的探讨奥曲肽与磷脂酰肌醇3激酶(PI3′K)通道抑制剂wortmannin联合用药对人胃癌细胞生长的影响及作用机制。方法2007年6月至8月在武汉大学人民医院消化实验室培养人胃癌细胞BGC-823,以单用浓度为40nmol/L的wortmannin,奥曲肽4个稀释度10-5、10-4、10-3及10-2g/L及它们各稀释度与wortmannin(40nmol/L)的混合培养液作用24h后噻唑蓝(MTT)比色实验检测细胞存活率,单用奥曲肽稀释度为10-3g/L和wortmannin以及二者的混和液作用12、24、36、48h检测细胞存活率,于24h检测细胞周期及各组p27基因信使核糖核苷酸(mRNA)和蛋白表达。结果单用奥曲肽组随浓度增加,抑制作用增强,浓度为10-3g/L时,抑制作用明显较对照组增强;单用wortmannin,有较强的肿瘤细胞抑制生长作用,且两者联合用药抑制作用更加明显;单用奥曲肽浓度组随时间的延长其抑制效率逐渐下降,与wortmannin共同作用时,未见细胞明显耐药;通过细胞周期分析,奥曲肽和wortmannin均能抑制肿瘤细胞使其生长停留在G1期,两者联合用药抑制作用更强;在联合用药组中p27蛋白表达明显升高,且随奥曲肽浓度增加表达增加;p27基因mRNA表达在各组无明显差异。结论奥曲肽和wortmannin对胃癌细胞BGC-823均有较强的抑制和杀伤作用,奥曲肽的抑制和毒性作用具有时间和浓度的依赖性,两者联合用药抑制和毒性效果更显著。PI3′K抑制剂wortmannin和奥曲肽均能使肿瘤细胞停止生长在G1期。Objective To investigate the effect and mechanism of octreotide with or without wortmannin on the growth of gastric carcinoma cell BGC-823. Methods From June to August of 2007, in Remin Hospital of Wuhan University, cells were exposed to octreotide for four dilution（ 10^-5,10^-4,10^-3,10^-2g/L） with or without wortmannin（ a special inhibitor of PI3 ＇K/Akt pathway, the dilution is 40 nmol/L）. And then the cytotoxicity was assessed by determining cell survival with MTT after 24 hours,with also were exposed to octreotide for the dilution of 10^-3 g/L with or without Wortmannin in 12 h, 24 h,36 h,48 h,then the cell survivral rate was accounted and the results were formulated in a table,and the cell cycle was checked-out with Flow Cytometer（FCM） ;the expression level of p27 gene and protein was determinated with RT-PCR and Western-bloting Test. Results The five groups within one reagent treating were compared with control group, and the results had difference, but the combination of two reagents group had significant difference（ P 〈 0.01 ）. Octreotide for dilution 10 -3 g/L alone had a time-dependent antiproliferation effect on BGC-823, but within wortmannin the result was changed. FCM revelead that octreotide and wortmannin could both inhibit the growth of gastric carcinoma cells,which were arrested in the period of time（ G1 ）. The check-out of p27 protein with Western-Bloting Test showed that the p27 protein expressed very strongly in the combination group, compared with Wort, Octe and control groups, having significant difference（P 〈0.01 ）. Conclusion Octreotide and wortmannin both have the effect of inhibiting and leading apoptosis to BGC-823 cells;octreotide has a dose-dependent and time-dependent antiproliferation effect on BGC-823 cells. But with wortmannin,the effect has significant difference. So octreotide and wortmannin both arrested cell growth at（ G1 ）phase.

关 键 词：磷脂酰肌醇3激酶通道 奥曲肽 WORTMANNIN P27蛋白 细胞增殖 

分 类 号：R5[医药卫生—内科学]