莪术醇对人胃癌细胞凋亡、MMP2、NO影响的初步

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胃癌在我国是最常见的恶性肿瘤之一,手术是其最有效的治疗方法,对于术后复发失去再次手术机会的及晚期胃癌患者,化疗仍是最有效的手段之一,但其疗效不尽人意,因为在消灭肿瘤细胞的同时,受其毒副作用影响而表现出较差的预后。因而寻求一种新的治疗方案迫在眉睫。肿瘤的靶向治疗基于分子水平,通过抑制相关蛋白或基因在信号转导通路、血管生成等途径发挥较强的抗肿瘤作用,且细胞毒性作

莪术醇对人胃癌细胞凋亡、MMP2、NO影响的

初步探讨

徐立春陈海燕文洁陶亚玲

扬州大学医学院肿瘤防治研究中心,江苏扬州225001

[摘要]目的初步探讨莪术醇(Curcumol)对人胃腺癌细胞(SGC-7901细胞)凋亡、基质金属蛋白酶(MMP2)、一氧化氮(NO)的影响。方法将莪术醇溶于无水乙醇中,初始浓度为9mg/mL,设实验1、2、3、4、5组,莪术醇的浓度分别是

7.5、15、30、60、120μg/mL,并以含1%无水乙醇的培养液为对照组。采用台盼蓝(Trypan blue)、噻唑蓝(MTT)法检测

SGC-7901细胞的活性及莪术醇对SGC-7901细胞抑制增殖的影响;用流式细胞仪(FACSCalibur)检测莪术醇对SGC-7901细胞的凋亡率;用蛋白质印迹法检测莪术醇作用于SGC-7901细胞后,其MMP2的表达情况;NO试剂盒检测莪术醇作用SGC-7901细胞后的细胞培养液上清的NO含量。结果30μg/mL的莪术醇在药物浓度与抑制肿瘤生长方面效果较佳,与对照组比较,其抑制肿瘤细胞增殖差异有统计学意义(P<0.05);莪术醇实验组能促进细胞凋亡,与对照组比较差异有统计学意义(P<0.05);莪术醇实验组作用细胞后,其MMP2蛋白表达明显下降,与对照组比较差异有统计学意义(P<0.05);实验研究表明,随着莪术醇实验组浓度的增加,其细胞培养液上清中的NO含量逐渐降低,与对照组比较差异有统计学意义(P<0.05)。结论30μg/mL的莪术醇浓度是最佳药效组,它能显著抑制SGC-7901细胞的增殖,明显促进肿瘤的细胞凋亡,显著抑制MMP2的表达,降低细胞培养液上清中的NO含量,从而发挥系列抗肿瘤作用。本研究从细胞分子水平探讨莪术醇治疗胃癌可能存在的部分机制,为临床应用莪术醇及其衍生物或结构类似物治疗胃癌提供理论与试验依据。

[关键词]莪术醇;SGC-7901;凋亡;基质金属蛋白酶;一氧化氮

[中图分类号]R000[文献标识码]A[文章编号]1673-7210(2012)12(a)-0018-04

Preliminary discussion on effect of curcumol in apoptosis,MMP2and NO in SGC-7901cells

XU Lichun CHEN Haiyan WEN Jie TAO Yaling

Cancer Prevention Research Center,Medical College of Yangzhou University,Jiangsu Province,Yangzhou225001,China [Abstract]Objective To discuss the effects of Curcumol on the apoptosis,MMP2and NO of human gastric cancer cells (SGC-7901cells).Methods Curcumol was dissolved in dehydrate alcohol and the initial concentration of Curcumol was9 mg/mL.Group1,group2,group3,group4and group5were set up as experimental groups.The Curcumol concentrations were set as follows:7.5,15,30,60,120μg/mL,control group contained1%dehydrate alcohol(dissolvent compare).The Trypan blueand method was used to analyze the cytoactive of SGC-7901;MTT assay was performed to study the inhibitory rate of SGC-7901cells proliferation;the metastasis of SGC-7901was described through the scratches experiment;the apoptosis rate were detected by FACSCalibur and the expression of Matrix metalloproteinase2(MMP2)were detected by Western Blot.The content of NO in cell culture medium was detected by NO kit.Results Curcumol had a good effect on drug concentration and restrain the tumor growth in30μg/mL,and compared with controls,it had significant difference in restrain the proliferation of tumor cell(P<0.05);Curcumol could promote the apoptosis,it showed significant difference when compared with controls(P<0.05);Curcumol could make the expression of MMP2protein descend to varying de-grees,it had significant difference when compared with controls(P<0.05);this research showed with the increasing of the concentration of curcumol,the content of the NO in the supernate gradually reduced,it had significant difference when compared with controls(P<0.05).Conclusion Curcumol in30μg/mL is the best efficacy group,it can significantly inhib-it the proliferation of SGC-7901cell,significantly promote the tumor cell apoptosis,significantly inhibit the expression of MMP2,reduce the content of NO in supernate,and express the series antitumor function.This study discusses the possible mechanism of curcumol cure the stomach cancer from cell and molecular level;provide the basis in theory and experiment for clinical application that Curcumol and its derivatives or structure analogues treat stomach cancer.

[Key words]Curcumol;SGC-7901;Apoptosis;MMP2;NO

[基金项目]江苏省医学卫生重点基金资助项目(编号:K200511)。

[作者简介]徐立春,研究员,教授,主要从事肿瘤防治基础与临床方面的

研究。

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