【作者】 李天望；

【导师】 凃晋文；

【作者基本信息】 湖北中医学院 ， 中医内科学， 2009， 博士

【摘要】 目的大肠癌是目前最常见恶性肿瘤之一,近年来我国的大肠癌的发病率呈上升趋势。祖国医学认为直肠癌的形成是湿热蕴结日久化毒下注,积聚于肠道凝结而成肿瘤。湿热下注、火热内蕴、结而为肿是病之标;正气不足、脾肾两亏为病之本。中医药治疗大肠癌是目前国内肿瘤治疗领域一大优势,大量实验及临床案例证明,中医药辅助治疗可以有效减轻放化疗副作用。我们在临床研究中,按照清热解毒、益气养阴、活血燥湿的治疗原则,用黄白抑瘤方口服及局部治疗该病取得了较好的临床疗效。在本实验中,我们拟采用人结直肠癌细胞株Colo320为研究对象,观察黄白抑瘤方在体外对Colo320细胞的抗肿瘤作用研究,探讨黄白抑瘤方对Colo320细胞增殖、凋亡,及其相关蛋白表达的影响,以明确黄白抑瘤方治疗结直肠癌的生物学基础。方法使用不同浓度黄白抑瘤方体外干预Colo320细胞。采用甲基噻唑MTT比色法和生长曲线测定不同浓度黄白抑瘤方对Colo320细胞的增殖的影响。采用流式细胞术检测不同浓度黄白抑瘤方对Colo320细胞周期和凋亡率的影响,并与对照组细胞的凋亡率比较,以显示黄白抑瘤方作用与细胞凋亡率之间的关系;Western blot检测不同浓度黄白抑瘤方处理后Colo320细胞的增殖细胞核抗原(Proliferating cell nuclear antigen,PCNA),以及凋亡相关蛋白Caspase3和PARP的表达。结果黄白抑瘤方作用Colo320细胞24h后,经MTT法检测,OD值明显小于对照组,显示不同浓度药物具有不同程度的肿瘤抑制率;黄白抑瘤方作用Colo320细胞48h后,与对照组相比,细胞周期分布发生明显改变,表现为S期细胞比例下降,G0/G1期和G2/M期细胞比例上升,凋亡率明显升高;黄白抑瘤方作用Colo320细胞48h后,与对照组相比,PCNA含量显著减少,而凋亡相关蛋白Caspase 3和PARP含量显著增加。结论黄白抑瘤方能抑制人直肠癌Colo320细胞增殖,并诱导促进细胞凋亡,从而达到抗肿瘤的作用。更多还原

【Abstract】 ObjectiveHuman rectal cancer is one of the most common malignant tumors. Recently,in our country,the incidence of rectal cancer is rising.In traditional Chinese medicine,the formation of rectal cancer is considered that damp-hot accumulates into toxin accumulating in intestinal,becoming a tumor. Damp-hot and fire-hot accumulating,which condense into a mass that is the Biao;Deficiency of vital Qi and deficiency of both spleen and kidney;that is the Ben? Currently,Chinese medicine treatment of rectal cancer is prevailing.Plenty of experimental and clinical cases have proved Chinese medicine can effectively reduce the toxicity of radiotherapy and chemotherapy. In clinical study,according to therapeutic rules of clearing away heat and toxic, reinforcing Qi and nourishing Yin,promoting blood and drying the damp,we used Huang Bai Yi Liu Fang to oral and local treatment and have achieved good clinical effect.For a better understanding of it’s roles in Cancer therapy, we adopted human rectal cancer cells Colo320 in this experiment to study it’s effect on Colo320 cell’s proliferation,apoptosis,and expression of related proteins.In order to define the biology basic of Huang Bai Yi Liu Fang on treating rectal cancer.MethodsDifferent concentrations of Huang Bai Yi Liu Fang was used to intervene Colo320 cells in vitro.MTT assay and growth curve were used to show the influence on Colo320 cell’s proliferation.Cell apoptosis and cell cycle distribution were determined by flow cytometry.The protein level of PCNA,PARP and caspase3 were examined by western blot analysis.ResultsOur data showed that Huang Bai Yi Liu Fang can inhibit the proliferation of Colo320 cells obviously;Compared with the control group,after intervened 48h,distribution of cell cycle have had significant changes in performance that display a sharp decline in the proportion of S phase and clear increase in GO / G1 phase and G2 / M phase’s cells,followed the apoptosis rate’s significantly increased;By 24h’s intervened of Huang Bai Yi Liu Fang, compared with the control group,PCNA had a significantly reduction, Caspase 3 and PARP increased sharply.ConclusionHuang Bai Yi Liu Fang can inhibit the growth of rectal cancer Colo320 cells through inducing apoptosis and cell cycle arrest,so as to achieve an anti-tumor role.更多还原