黄白抑瘤方对人直肠癌细胞增殖和凋亡的影响
|
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。
Objective
Human 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.
Methods
Different 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.
Results
Our 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.
Conclusion
Huang 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.
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。
Objective
Human 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.
Methods
Different 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.
Results
Our 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.
Conclusion
Huang 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.
引文
1.Parkin DM,Bray F,Gerlay J,et al.Global cancer statistics,2002.CA:a cancer journal for clinicians 2005;55:74-108.
2.International Muhicentre Pooled Analysis of Colon Cancer Tri.Als(IMPACT)investigators.Eficacy of adjuvant fluorouracil and folinic acid in colon cancer.Lancet 1995;345:939.
3.Doekhie FS,Morreau H,de Bock GH,et al.Sialyl Lewis X Expression and Lymphatic Microvessel Density in Primary Tumors of Node-negative Colorectal Cancer Patients Predict Disease Recurrence.Cancer Microenviron.2008 Dec;1(1):141-51.Epub 2008 Aug 6.
4.Gerard JP,Francois E,Deustch E,et al.Neoadjuvant approach for non-metastatic rectal cancer combining radiotherapy and chemotherapy.Biotargeted drugs.]Bull Cancer.2009 Mar 1;96(3):321-328.French.
5.Apati A,Janossy J,Brozik A,et al.Calcium induces cell survival and proliferation through the activation of the MAPK pathway in a human hormone-dependent leukemia cell line,TF-l.J Biol Chem.2003 Mar 14;278(11):9235-43.
6.Schettini J,Mukherjee P.Physiological role of plasmacytoid dendritic cells and their potential use in cancer immunity.Clin Dev Immunol.2008;2008:106321.Epub 2009 Jan 26.
7.Kuribayashi K,Krigsfeld G,Wang W,et al.TNFSFlO(TRAIL),a p53 target gene that mediates p53-dependent cell death.Cancer Biol Ther.2008 Dec;7(12):2034-8.
8.Casado E,De Castro J,Belda-Iniesta C,et al.Molecular markers in colorectal cancer:genetic bases for a customised treatment.Clin Transl Oncol.2007Sep;9(9):549-54.Review.
9.Kimitohi I,Ynichi Q,Tashyuki S,et al.Vassoular endothelial growth factor expression in primary eso-phaus squamous cellcarcinoma association mith arngiogenesis and tumor progress[J].Cancer Res,1997,(79):206
10.李博,李瑞汉,高冬梅.中医药抗肿瘤机理研究概况[J].实用中医内科杂志,2005,19(5):403-405.
11.黄泰康.常用中药成分与药理手册[M].北京:中国医药科技出版社,1994:228-229.
12.蔡永敏.最新中药药理与临床应用[M].北京:华厦出版社,1999:125-129.
13.姜晓峰,甄永苏.大黄逆转肿瘤细胞多药抗药性作用[J].药学学报,1999,34(3):164-165.
14.Huang Q,Shen HM,Ong CN.Emodin inhibits tumor cell migratlthrough suppression of the phosphatidylinos 3-kinase-Cdc42/Racl pathway[J].Cell Mol Life Sci,2005,62(10):116.
15.Huang Q,Shen HM,Ong CN.Inhibitory efect of emodin on tumor invasion through suppresion of activator protein-1 and nuclear fact kappaB[J].Biochem pharmacol,2004,68(2):361.
16.章荣华,张仲苗.白头翁对二甲基肼诱发小鼠大肠癌的防治作用和机理研究.中药药理与临床,1999年,15(5):62-64.
17.黄芳,李娟.西南白头翁不同提取物的抗肿瘤作用研究.湖北中医学院学报,2008,10(3):12-13.
18 蔡鹰,余国祥.白头翁抗肿瘤机理研究.云南中医中药杂志,2001,22(5):32-33.
19 蔡鹰,等.中草药.1999.(30)13:92.
20.吴歌,杨世林.人参皂苷单体Rh2诱导小鼠前胃癌细胞系凋亡的分子机制.中国药理学通报,2008,24(1):21-23.
21.Liu WK,Xu SX,Che CT.Anti-pmliferative effect of ginseng sapoins on human prostate cancer cell line[J].Life Sci,2000,67(11):1297-1360.
22.Kim S E,LeYH,Park JH。et al.Gi nsenoside-Rs3,a new diol-type ginseng saponin,selectively elevates protein levels of p53 and p21 WAF1leading to induction of apoptosis in SK-HEP-1 cels[J].Anticancer Res,1999,19(1A):487-491
23.钱伯文.抗癌中药的临床效用.上海:上海翻译出版公司,1987.10.
24.常敏毅.抗癌本草.长沙:湖南科学技术出版社,1987.105.
25.刘春安,彭明.抗癌中草药大辞典.武汉:湖北科学技术出版社,1994.392.
26.张锡纯.医学衷中参西录.中册.石家庄:河北科学技术出版社,1985.90.
27.杨今祥.抗癌中草药制剂.北京:人民卫生出版社,1991.88.
28.何萍,李帅.半夏化学成分的研究.中国中药杂志,2005,30(9):671-674.
29.大原直子.半夏免疫系统激活多糖的分离和性状[J].国外医学·中医中药分册,1993,15(3):56.
30.国家中医药管理局《中华本草》编委会.中华本草(8).上海:上海科学技术出版社,1999.626.
31.3]国家中医药管理局《中华本草》编委会.中华本草(8).上海:上海科学技术出版社,1999.626
32..施广霞,于丽华,刘金友,等.β榄香烯抗肿瘤作用的实验研究:β 榄香烯体外对k15白血病细胞直接作用的实验研究[J].大连医学院学报,1994,16(2):137.
33.杨骅,王仙平,郁琳琳,等.榄香烯抗癌作用与诱发肿瘤细胞凋亡[J].中华肿瘤杂志,1996,18(3):169.
34.吴万垠,罗云坚,程剑华,等.莪术油对小鼠肝癌细胞DNA作用的图像分析[J].中西医结合肝病杂志,1999,9(1):18.
35.崔秀云,李德山.β榄香烯对RNA聚合酶活性的抑制及与DNA的结合[J].中国药理学报,1991,7(3):228.
36.卜长武,王东荣,郭文来,等.莪术对小白鼠免疫功能的影响[J].中国中医药信息杂志,1997,4(2):29.
37.钱振超,王大庆,金成刚,等.瘤苗特异性主动免疫治疗及其机制的实验研究[J].肿瘤生物治疗杂志,1999,2(2):125.
38.陈红,邓德宁等.莪术醇及莪术醇磷酸酯单钠对Ehrlich腹水癌细胞电位和钾离子通道的影响[J].首都医学院学报,1989,10(1):23.
39.于冬青,邓华聪.姜黄素的药理研究进展[J].山东医药,2005,45(2):72-73.
40.International Muhicentre Pooled Analysis of Colon Cancer Tri.Als (IMPACT)investigators.Eficacy of adjuvant fluorouracil and folinic acid in colon cancer.Lancet 1995;345:939.
41.Carmichael J,DeGraffWG,GazdarAF,et al.Evaluation of a tetrazoliumbased semiautomated colorimetric assay:assessment of chemosensitivity testing[J].Cancer Res,1987,47(4):936.
42.Lee MS,Sun MT,Pang ST,et al.TH Evaluation of differentially expressed genes by shear stress in human osteoarthritic chondrocytes in vitro.Chang Gung Med J.2009 Jan-Feb;32(1):42-50.
43.Gao CY,Zeienka PS.Cyclins,cyclin-dependent kinases and differentiation.Bioessays,1997,19:307-15.
44.Kim IS,Song JK,Song YM,et al.Novel Effect of Biphasic Electric Current on In Vitro Osteogenesis and Cytokine Production in Human Mesenchymal Stromal Cells.Tissue Eng Part A.2009 Jan 21.
45.Nishitani H,Lygerou Z.Contral of DNA replication licensing in a cellcycle.Genes Cells,2002;7(6):523-34.
46.Kanematsu T,Maehara Y,Matsumata T,et al.Human hepatocellular carcinoma sensitivity to antitumor drugs assayed using the succinate dehydrogenase inhibition test[J].Oncology,1991,48(1):34～38.
47.Tye BK.Initiating DNA synthesis:from recruiting to activating theMCM complex[J].J Cell SCi,2001;(114):1447-54.
48.Hartwell LH,Kastan KB.Cell cycle control and cancer.Science 1994;266:1821-1036.
49.Hunter T,Pines J.Cyclins and cancer.Cell 1991;66:1071-1074.
50.Marx J.How cell cycle toward cancer.Science 1994;263:319-321.
51.King KL,Cidlowwski JA.Cell cycle regulation and apoptosis.Annu Res Physiol 1998;60;601-617.
52.Cluman BE,Roberts JM.Cell cycle and cancer.JNCI 1995;87:1499-1501.
53.Schwartz LM,O sbo rne BA.P rogrammed cell death,apoptosis and k iller genes.Immunol Today,1993,14(12):582-590.
54.Villegas I,Sanchez-Fidalgo S,Alarcon de la Lastra C.New mechanisms and therapeutic potential of curcumin for colorectal cancer.Mol Nutr Food Res.2008 Sep;52(9):1040-61.
55.Johnson K,Tan M,Sutterlin C.Centrosome abnormalities during a Chlamydia trachomatis infection are caused by dysregulation of the normal duplication pathway.Cell Microbiol.2009 Mar 12.
56.Gajecka M,Obremski K,Jakimiuk E,et al.Histopathological examination of ovaries in bitches after experimental zearalenone mycotoxicosis.Pol J Vet Sci.2008;11(4):363-6.
57.Cao Yunxin,Jin Weilin,Jin Boquan,et al.A new method specified for detecting apoptosis and cell cycles.Chinese journal of medical research.2003,12-1065-02.
58.Anjomshoaa A,Lin YH,Black MA,et al.Reduced expression of a gene proliferation signature is associated with enhanced malignancy in colon cancer.Br J Cancer.2008 Sep 16;99(6):966-73.
59.Anjomshoaa A,Lin YH,Black MA,et al.Reduced expression of a gene proliferation signature is associated with enhanced malignancy in colon cancer.Br J Cancer.2008 Sep 16;99(6):966-73.
60.Bravo R,Frank R,Blundell PA et al.Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta.Nature 1987;332(6112):515-7.
61.Prelich G,Kostura M,Marshak D R et al.The cell cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro.Nature 1987;326(6112):471-5.
62.Melnick M,Chen H,Min Zhou Y,.et al.The functional genomic response of developing embryonic submandibular glands to NF-kappa B inhibition.BMC Dev Biol.2001;1:15.Epub 2001 Oct 25.
63.Porter A G,Janicke R U.Emerging roles of caspase-3 in apotosis.Cell Death Differ,1999;6(2):99.
64.Rodriguez LC,AlpHonse,Broquet P,et al.Temporal relationships between ceramide.caspase activation and mitochondrial dysfunction in cell lines with arying sensitivity to anti—Fas-—induced apoptosis.Biochem J,2001,357(Pt)2:407-16.
65.Tan KY,Liu CB,Chen AH,et al.The role of traditional Chinese medicine in colorectal cancer treatment.Tech Coloproctol.2008 Mar;12(1):1-6;discussion 6.Epub 2008 May 30.
66.Prelich G,Kostura M,Marshak D R et al.The cell cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro.Nature 1987;326(6112):471-5.
67.Lee HZ,Wu CH,Chang SPlRelease of nucleophosmin from the nucleus:Involvement in aloe-emodin-induced human lung non small carcinoma cell apoptosis[J]1 Int J Cancer,2005(113):971-9761.
68.Prelich G,Kostura M,Marshak D R et.al.The cell cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro.Nature 1987;326(6112):471-5.
69.Apati A,Janossy J,Brozik A,et al.Calcium induces cell survival and proliferation through the activation of the MAPK pathway in a human hormone-dependent leukemia cell line,TF-l.J Biol Chem.2003 Mar 14;278(11):9235-43.
70.Lutsenko MT.Computer assessment of an apoptotic process dynamics in the nuclei of placental villi syncytiotrophoblast in puerperas after the outburst of herpes virus infection.Morfologiia.2008;134(6):79-83.Russian.
1.Mckay JA,Douglas JJ,Ross VG,et al.Analysis of keycell-cy cle checkpoint proteins in colorectal tumoursEJ-].Pathol,2002,196(4):386-393.
2.NasmythK,HunterT.cell cycle:Damsand sluices、Nature,1993;366(6465):634-635.
3.Horrwe11LH,Kastan MB.Cell cycle control and cancer.Science.1994:266(5198):1821-1828.
4.陈小贺、孟文格等.细胞周期相关蛋白CyclinD1、CDK4、p16在大肠癌中的表达及其意义。实用肿瘤学杂志[J];2008.22(4)317-321.
5.Dierendonck JH,Wijsman JH,Keijzer R,et al.Cell-cycle-related staining patterns of antiproliferating cell nuclear ant igen monoclonal antibodies.Comparison with Brd Urd labeling an d Ki-67 staining[J′].Am J Pathol,1991,138(5):1165.
6.胡庆军,周茜.表皮生长因子受体在大肠癌中的表达及与细胞增殖状态的关系.中国煤炭工业医学杂志[J];2007.10(8):926-927.
7.杨宾、付强、王亚娟、唐琢.芦荟大黄素对人结肠癌细胞SW480增殖周期及凋亡的影响.山西中医[J].2008.24(6)48-50.
8.宋寄春,谭诗云,陈明锴。半枝莲对人大肠癌细胞增殖抑制及诱导凋亡的作用[J].成宁学院学报.2007,21(2):601-603.
9.Macian F.NFAT proteins:key regulators ofT-cel development a nd function.Nat Rev linmunol,2005;5:472-484.
10.李小军、 张卫军、田伏洲、尹致良、雷士伟.槲皮素对大肠癌细胞SW116增殖的影响.成都军区医院学报[J].2001.3(2):1-2.
11.Cai Y,Zhang L,Li N,et al.Shuo.Cai.A Study on ultrastr uctural localization ang activity of the lymphoprotease of huma n lymph node.US-China Med Sci,2005,2(4):47-50.
12.张丽红,陈晓琳,王旭,蔡玉文.益肠泰对人大肠癌细胞株CCL229细胞增殖和酶活性的抑制作用.解剖科学进展[J].2007.13(2):155-157.
1.黎明红,姚震.妇科恶性肿瘤与细胞凋亡[J].医学综述,2002,8(1):5271.
2.Kerr J FR,Wyllie AH,Currie AR.Apoptosis,a basic biological phenomenon with wider implications in tissue kinetics[J].Br J Cancer,1972,26:2392245.
3.杨宾,付强,等。芦荟大黄素对人结肠癌细胞SW480增殖周期及凋亡的影响[J].山西中医.2008,24(6):48-50.
4.宋寄春,谭诗云,等.半枝莲对人大肠癌细胞增殖抑制及诱导凋亡的作用[J].成宁学院学报.2007,21(2):601-603.
5.刘得育,罗曼等.蛇葡萄素抗肿瘤作用研究[J].癌症,2001,20(12):1372-1375.
6.Vogelstein B.Genetic alteration during colorectal tumor development[J].N Engl J Med,1988,319:525.
7.司维柯,高利宏,等.苦参碱诱导人肝癌细胞分化、凋亡时对细胞周期调节因子的调控[J].癌症,2001,20(8):848-851.
8.徐群英.抗癌中药药理研究概况[J].时珍国医国药.2001,12(2)
9.庄肾韩,耿宝琴,等.白头翁抗肿瘤作用实验研究[J].实用肿瘤杂志,1999,14(2):94.
10.钱丽娟,许沈华,等.鳖甲浸出液对大肠癌细胞(HR-8348)抑制作用的形态学观察[J].癌症,1997,16(3):175.
11.蒋英丽,宋今丹.新疆紫草素诱导人大肠癌细胞的凋亡[J].癌症,2001,20(12):13552-13581.
1.叶任高.内科学,北京:人民卫生出版社,2000,5:437.
2.刘嘉湘.现代中医药应用与研究大系·肿瘤科分册.上海:上海中医药大学出版社,1996,1:189-190.
3.高虹,朱晏伟.刘嘉湘教授治大肠癌.实用中西医结合杂志,1996;9(10):630-631.
4.陈培丰.单纯中医药治疗晚期直肠癌18例.陕西中医,1995;16(1):12.
5.张新,孙华,等.孙桂芝治疗大肠癌经验.山东中医杂志,1998;17(4):173-174.
6.许环应,李财宝,等.大肠癌术后加用扶正解毒中绪的疗效观察.上海中医药杂志,1996,(2):12.
7.钱伯文.上海中医药大学中医学家专集,北京:人民卫生出版社,1999:58-59.
8.曹洋,刘展华,等.陈锐深教授治疗大肠癌的经验[J].中医药学刊,2005,23(10):1750-1751.
9.张恩欣.周岱翰论治大肠癌经验[J].实用中医药杂志,2006,22(9):562.
10.王小宁,霍介格.中医治疗大肠癌的思路与方法探讨中国中医[J].基础医学杂志,2007,13(9):681-682.
11.丁金芳,黄云胜,等.施志明治疗大肠癌经验举要[J].上海中医药杂志,2007,41(5):43-44.
12.田劭丹,李忠.大肠癌的中西医结合诊治[J].中国临床医生杂志,2007,35(4):26-29.
13.贾小强,邱辉忠,等.大肠癌辨证分型与肿瘤浸润转移相关性的前瞻性研究[J].中华中医药杂志,2005,20(6):344-346.
14.魏星,裘华森,周济春.血瘀证辨证对结直肠癌淋巴结转移的预测作用[J].浙江临床医学,2007,9(10):1392-1393.
15.顾缨,韩颖盈,等.胃肠安治疗大肠癌临床疗效分析[J].辽宁中医药大学学报,2006,8(5):5-6.
16.秦银忠,黄晓峰,李志刚.清藏固本汤配合化疗治疗中晚期大肠癌疗效观察[J].现代中西医结合杂志,2006,15(24):3377-3378.
17.赵文硕,张青,等.加味升血汤配合化疗治疗中晚期大肠癌的临床研究[J].国际中医中药杂志,2006,28(5):306-308.
18.蒋益兰,潘博,仇湘中.健脾消癌饮配合化疗治疗大肠癌术后40例总结[J].湖南中医杂志,2001,17(5):9-10.
19.王振飞.大肠癌术后化疗配合艾迪注射液临床观察[J].福建中医药,2007,38(4):9-10.
20.尤建良,周留勇.扶正和胃合剂治疗大肠癌48例[J].药学与临床研究 2007,15(1):60-62.
21.杨彦,刘碧清,杜秋霞.肠复康胶囊治疗中晚期大肠癌55例临床观察[J].江苏中医药,2005,26(1):18-20.
22.金哲秀.针灸两步法治疗大肠癌27例临床分析.上海中医药杂志2003,37:48-49.
23.穆立新,姜军作,范杰华.针灸治疗大肠癌术后30例疗效观察.大连大学学报,2006,27:100-106.
24.黄兆明.中药灌肠综合治疗癌性肠梗阻临床观察[J].实用中西医结合临床,2003,3(3):36.
25.郑玉玲,王新杰.肠达顺灌肠液治疗湿热蕴结型大肠癌的临床研究[J].河南中医,2002,22(1):12.
2.International Muhicentre Pooled Analysis of Colon Cancer Tri.Als(IMPACT)investigators.Eficacy of adjuvant fluorouracil and folinic acid in colon cancer.Lancet 1995;345:939.
3.Doekhie FS,Morreau H,de Bock GH,et al.Sialyl Lewis X Expression and Lymphatic Microvessel Density in Primary Tumors of Node-negative Colorectal Cancer Patients Predict Disease Recurrence.Cancer Microenviron.2008 Dec;1(1):141-51.Epub 2008 Aug 6.
4.Gerard JP,Francois E,Deustch E,et al.Neoadjuvant approach for non-metastatic rectal cancer combining radiotherapy and chemotherapy.Biotargeted drugs.]Bull Cancer.2009 Mar 1;96(3):321-328.French.
5.Apati A,Janossy J,Brozik A,et al.Calcium induces cell survival and proliferation through the activation of the MAPK pathway in a human hormone-dependent leukemia cell line,TF-l.J Biol Chem.2003 Mar 14;278(11):9235-43.
6.Schettini J,Mukherjee P.Physiological role of plasmacytoid dendritic cells and their potential use in cancer immunity.Clin Dev Immunol.2008;2008:106321.Epub 2009 Jan 26.
7.Kuribayashi K,Krigsfeld G,Wang W,et al.TNFSFlO(TRAIL),a p53 target gene that mediates p53-dependent cell death.Cancer Biol Ther.2008 Dec;7(12):2034-8.
8.Casado E,De Castro J,Belda-Iniesta C,et al.Molecular markers in colorectal cancer:genetic bases for a customised treatment.Clin Transl Oncol.2007Sep;9(9):549-54.Review.
9.Kimitohi I,Ynichi Q,Tashyuki S,et al.Vassoular endothelial growth factor expression in primary eso-phaus squamous cellcarcinoma association mith arngiogenesis and tumor progress[J].Cancer Res,1997,(79):206
10.李博,李瑞汉,高冬梅.中医药抗肿瘤机理研究概况[J].实用中医内科杂志,2005,19(5):403-405.
11.黄泰康.常用中药成分与药理手册[M].北京:中国医药科技出版社,1994:228-229.
12.蔡永敏.最新中药药理与临床应用[M].北京:华厦出版社,1999:125-129.
13.姜晓峰,甄永苏.大黄逆转肿瘤细胞多药抗药性作用[J].药学学报,1999,34(3):164-165.
14.Huang Q,Shen HM,Ong CN.Emodin inhibits tumor cell migratlthrough suppression of the phosphatidylinos 3-kinase-Cdc42/Racl pathway[J].Cell Mol Life Sci,2005,62(10):116.
15.Huang Q,Shen HM,Ong CN.Inhibitory efect of emodin on tumor invasion through suppresion of activator protein-1 and nuclear fact kappaB[J].Biochem pharmacol,2004,68(2):361.
16.章荣华,张仲苗.白头翁对二甲基肼诱发小鼠大肠癌的防治作用和机理研究.中药药理与临床,1999年,15(5):62-64.
17.黄芳,李娟.西南白头翁不同提取物的抗肿瘤作用研究.湖北中医学院学报,2008,10(3):12-13.
18 蔡鹰,余国祥.白头翁抗肿瘤机理研究.云南中医中药杂志,2001,22(5):32-33.
19 蔡鹰,等.中草药.1999.(30)13:92.
20.吴歌,杨世林.人参皂苷单体Rh2诱导小鼠前胃癌细胞系凋亡的分子机制.中国药理学通报,2008,24(1):21-23.
21.Liu WK,Xu SX,Che CT.Anti-pmliferative effect of ginseng sapoins on human prostate cancer cell line[J].Life Sci,2000,67(11):1297-1360.
22.Kim S E,LeYH,Park JH。et al.Gi nsenoside-Rs3,a new diol-type ginseng saponin,selectively elevates protein levels of p53 and p21 WAF1leading to induction of apoptosis in SK-HEP-1 cels[J].Anticancer Res,1999,19(1A):487-491
23.钱伯文.抗癌中药的临床效用.上海:上海翻译出版公司,1987.10.
24.常敏毅.抗癌本草.长沙:湖南科学技术出版社,1987.105.
25.刘春安,彭明.抗癌中草药大辞典.武汉:湖北科学技术出版社,1994.392.
26.张锡纯.医学衷中参西录.中册.石家庄:河北科学技术出版社,1985.90.
27.杨今祥.抗癌中草药制剂.北京:人民卫生出版社,1991.88.
28.何萍,李帅.半夏化学成分的研究.中国中药杂志,2005,30(9):671-674.
29.大原直子.半夏免疫系统激活多糖的分离和性状[J].国外医学·中医中药分册,1993,15(3):56.
30.国家中医药管理局《中华本草》编委会.中华本草(8).上海:上海科学技术出版社,1999.626.
31.3]国家中医药管理局《中华本草》编委会.中华本草(8).上海:上海科学技术出版社,1999.626
32..施广霞,于丽华,刘金友,等.β榄香烯抗肿瘤作用的实验研究:β 榄香烯体外对k15白血病细胞直接作用的实验研究[J].大连医学院学报,1994,16(2):137.
33.杨骅,王仙平,郁琳琳,等.榄香烯抗癌作用与诱发肿瘤细胞凋亡[J].中华肿瘤杂志,1996,18(3):169.
34.吴万垠,罗云坚,程剑华,等.莪术油对小鼠肝癌细胞DNA作用的图像分析[J].中西医结合肝病杂志,1999,9(1):18.
35.崔秀云,李德山.β榄香烯对RNA聚合酶活性的抑制及与DNA的结合[J].中国药理学报,1991,7(3):228.
36.卜长武,王东荣,郭文来,等.莪术对小白鼠免疫功能的影响[J].中国中医药信息杂志,1997,4(2):29.
37.钱振超,王大庆,金成刚,等.瘤苗特异性主动免疫治疗及其机制的实验研究[J].肿瘤生物治疗杂志,1999,2(2):125.
38.陈红,邓德宁等.莪术醇及莪术醇磷酸酯单钠对Ehrlich腹水癌细胞电位和钾离子通道的影响[J].首都医学院学报,1989,10(1):23.
39.于冬青,邓华聪.姜黄素的药理研究进展[J].山东医药,2005,45(2):72-73.
40.International Muhicentre Pooled Analysis of Colon Cancer Tri.Als (IMPACT)investigators.Eficacy of adjuvant fluorouracil and folinic acid in colon cancer.Lancet 1995;345:939.
41.Carmichael J,DeGraffWG,GazdarAF,et al.Evaluation of a tetrazoliumbased semiautomated colorimetric assay:assessment of chemosensitivity testing[J].Cancer Res,1987,47(4):936.
42.Lee MS,Sun MT,Pang ST,et al.TH Evaluation of differentially expressed genes by shear stress in human osteoarthritic chondrocytes in vitro.Chang Gung Med J.2009 Jan-Feb;32(1):42-50.
43.Gao CY,Zeienka PS.Cyclins,cyclin-dependent kinases and differentiation.Bioessays,1997,19:307-15.
44.Kim IS,Song JK,Song YM,et al.Novel Effect of Biphasic Electric Current on In Vitro Osteogenesis and Cytokine Production in Human Mesenchymal Stromal Cells.Tissue Eng Part A.2009 Jan 21.
45.Nishitani H,Lygerou Z.Contral of DNA replication licensing in a cellcycle.Genes Cells,2002;7(6):523-34.
46.Kanematsu T,Maehara Y,Matsumata T,et al.Human hepatocellular carcinoma sensitivity to antitumor drugs assayed using the succinate dehydrogenase inhibition test[J].Oncology,1991,48(1):34～38.
47.Tye BK.Initiating DNA synthesis:from recruiting to activating theMCM complex[J].J Cell SCi,2001;(114):1447-54.
48.Hartwell LH,Kastan KB.Cell cycle control and cancer.Science 1994;266:1821-1036.
49.Hunter T,Pines J.Cyclins and cancer.Cell 1991;66:1071-1074.
50.Marx J.How cell cycle toward cancer.Science 1994;263:319-321.
51.King KL,Cidlowwski JA.Cell cycle regulation and apoptosis.Annu Res Physiol 1998;60;601-617.
52.Cluman BE,Roberts JM.Cell cycle and cancer.JNCI 1995;87:1499-1501.
53.Schwartz LM,O sbo rne BA.P rogrammed cell death,apoptosis and k iller genes.Immunol Today,1993,14(12):582-590.
54.Villegas I,Sanchez-Fidalgo S,Alarcon de la Lastra C.New mechanisms and therapeutic potential of curcumin for colorectal cancer.Mol Nutr Food Res.2008 Sep;52(9):1040-61.
55.Johnson K,Tan M,Sutterlin C.Centrosome abnormalities during a Chlamydia trachomatis infection are caused by dysregulation of the normal duplication pathway.Cell Microbiol.2009 Mar 12.
56.Gajecka M,Obremski K,Jakimiuk E,et al.Histopathological examination of ovaries in bitches after experimental zearalenone mycotoxicosis.Pol J Vet Sci.2008;11(4):363-6.
57.Cao Yunxin,Jin Weilin,Jin Boquan,et al.A new method specified for detecting apoptosis and cell cycles.Chinese journal of medical research.2003,12-1065-02.
58.Anjomshoaa A,Lin YH,Black MA,et al.Reduced expression of a gene proliferation signature is associated with enhanced malignancy in colon cancer.Br J Cancer.2008 Sep 16;99(6):966-73.
59.Anjomshoaa A,Lin YH,Black MA,et al.Reduced expression of a gene proliferation signature is associated with enhanced malignancy in colon cancer.Br J Cancer.2008 Sep 16;99(6):966-73.
60.Bravo R,Frank R,Blundell PA et al.Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta.Nature 1987;332(6112):515-7.
61.Prelich G,Kostura M,Marshak D R et al.The cell cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro.Nature 1987;326(6112):471-5.
62.Melnick M,Chen H,Min Zhou Y,.et al.The functional genomic response of developing embryonic submandibular glands to NF-kappa B inhibition.BMC Dev Biol.2001;1:15.Epub 2001 Oct 25.
63.Porter A G,Janicke R U.Emerging roles of caspase-3 in apotosis.Cell Death Differ,1999;6(2):99.
64.Rodriguez LC,AlpHonse,Broquet P,et al.Temporal relationships between ceramide.caspase activation and mitochondrial dysfunction in cell lines with arying sensitivity to anti—Fas-—induced apoptosis.Biochem J,2001,357(Pt)2:407-16.
65.Tan KY,Liu CB,Chen AH,et al.The role of traditional Chinese medicine in colorectal cancer treatment.Tech Coloproctol.2008 Mar;12(1):1-6;discussion 6.Epub 2008 May 30.
66.Prelich G,Kostura M,Marshak D R et al.The cell cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro.Nature 1987;326(6112):471-5.
67.Lee HZ,Wu CH,Chang SPlRelease of nucleophosmin from the nucleus:Involvement in aloe-emodin-induced human lung non small carcinoma cell apoptosis[J]1 Int J Cancer,2005(113):971-9761.
68.Prelich G,Kostura M,Marshak D R et.al.The cell cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro.Nature 1987;326(6112):471-5.
69.Apati A,Janossy J,Brozik A,et al.Calcium induces cell survival and proliferation through the activation of the MAPK pathway in a human hormone-dependent leukemia cell line,TF-l.J Biol Chem.2003 Mar 14;278(11):9235-43.
70.Lutsenko MT.Computer assessment of an apoptotic process dynamics in the nuclei of placental villi syncytiotrophoblast in puerperas after the outburst of herpes virus infection.Morfologiia.2008;134(6):79-83.Russian.
1.Mckay JA,Douglas JJ,Ross VG,et al.Analysis of keycell-cy cle checkpoint proteins in colorectal tumoursEJ-].Pathol,2002,196(4):386-393.
2.NasmythK,HunterT.cell cycle:Damsand sluices、Nature,1993;366(6465):634-635.
3.Horrwe11LH,Kastan MB.Cell cycle control and cancer.Science.1994:266(5198):1821-1828.
4.陈小贺、孟文格等.细胞周期相关蛋白CyclinD1、CDK4、p16在大肠癌中的表达及其意义。实用肿瘤学杂志[J];2008.22(4)317-321.
5.Dierendonck JH,Wijsman JH,Keijzer R,et al.Cell-cycle-related staining patterns of antiproliferating cell nuclear ant igen monoclonal antibodies.Comparison with Brd Urd labeling an d Ki-67 staining[J′].Am J Pathol,1991,138(5):1165.
6.胡庆军,周茜.表皮生长因子受体在大肠癌中的表达及与细胞增殖状态的关系.中国煤炭工业医学杂志[J];2007.10(8):926-927.
7.杨宾、付强、王亚娟、唐琢.芦荟大黄素对人结肠癌细胞SW480增殖周期及凋亡的影响.山西中医[J].2008.24(6)48-50.
8.宋寄春,谭诗云,陈明锴。半枝莲对人大肠癌细胞增殖抑制及诱导凋亡的作用[J].成宁学院学报.2007,21(2):601-603.
9.Macian F.NFAT proteins:key regulators ofT-cel development a nd function.Nat Rev linmunol,2005;5:472-484.
10.李小军、 张卫军、田伏洲、尹致良、雷士伟.槲皮素对大肠癌细胞SW116增殖的影响.成都军区医院学报[J].2001.3(2):1-2.
11.Cai Y,Zhang L,Li N,et al.Shuo.Cai.A Study on ultrastr uctural localization ang activity of the lymphoprotease of huma n lymph node.US-China Med Sci,2005,2(4):47-50.
12.张丽红,陈晓琳,王旭,蔡玉文.益肠泰对人大肠癌细胞株CCL229细胞增殖和酶活性的抑制作用.解剖科学进展[J].2007.13(2):155-157.
1.黎明红,姚震.妇科恶性肿瘤与细胞凋亡[J].医学综述,2002,8(1):5271.
2.Kerr J FR,Wyllie AH,Currie AR.Apoptosis,a basic biological phenomenon with wider implications in tissue kinetics[J].Br J Cancer,1972,26:2392245.
3.杨宾,付强,等。芦荟大黄素对人结肠癌细胞SW480增殖周期及凋亡的影响[J].山西中医.2008,24(6):48-50.
4.宋寄春,谭诗云,等.半枝莲对人大肠癌细胞增殖抑制及诱导凋亡的作用[J].成宁学院学报.2007,21(2):601-603.
5.刘得育,罗曼等.蛇葡萄素抗肿瘤作用研究[J].癌症,2001,20(12):1372-1375.
6.Vogelstein B.Genetic alteration during colorectal tumor development[J].N Engl J Med,1988,319:525.
7.司维柯,高利宏,等.苦参碱诱导人肝癌细胞分化、凋亡时对细胞周期调节因子的调控[J].癌症,2001,20(8):848-851.
8.徐群英.抗癌中药药理研究概况[J].时珍国医国药.2001,12(2)
9.庄肾韩,耿宝琴,等.白头翁抗肿瘤作用实验研究[J].实用肿瘤杂志,1999,14(2):94.
10.钱丽娟,许沈华,等.鳖甲浸出液对大肠癌细胞(HR-8348)抑制作用的形态学观察[J].癌症,1997,16(3):175.
11.蒋英丽,宋今丹.新疆紫草素诱导人大肠癌细胞的凋亡[J].癌症,2001,20(12):13552-13581.
1.叶任高.内科学,北京:人民卫生出版社,2000,5:437.
2.刘嘉湘.现代中医药应用与研究大系·肿瘤科分册.上海:上海中医药大学出版社,1996,1:189-190.
3.高虹,朱晏伟.刘嘉湘教授治大肠癌.实用中西医结合杂志,1996;9(10):630-631.
4.陈培丰.单纯中医药治疗晚期直肠癌18例.陕西中医,1995;16(1):12.
5.张新,孙华,等.孙桂芝治疗大肠癌经验.山东中医杂志,1998;17(4):173-174.
6.许环应,李财宝,等.大肠癌术后加用扶正解毒中绪的疗效观察.上海中医药杂志,1996,(2):12.
7.钱伯文.上海中医药大学中医学家专集,北京:人民卫生出版社,1999:58-59.
8.曹洋,刘展华,等.陈锐深教授治疗大肠癌的经验[J].中医药学刊,2005,23(10):1750-1751.
9.张恩欣.周岱翰论治大肠癌经验[J].实用中医药杂志,2006,22(9):562.
10.王小宁,霍介格.中医治疗大肠癌的思路与方法探讨中国中医[J].基础医学杂志,2007,13(9):681-682.
11.丁金芳,黄云胜,等.施志明治疗大肠癌经验举要[J].上海中医药杂志,2007,41(5):43-44.
12.田劭丹,李忠.大肠癌的中西医结合诊治[J].中国临床医生杂志,2007,35(4):26-29.
13.贾小强,邱辉忠,等.大肠癌辨证分型与肿瘤浸润转移相关性的前瞻性研究[J].中华中医药杂志,2005,20(6):344-346.
14.魏星,裘华森,周济春.血瘀证辨证对结直肠癌淋巴结转移的预测作用[J].浙江临床医学,2007,9(10):1392-1393.
15.顾缨,韩颖盈,等.胃肠安治疗大肠癌临床疗效分析[J].辽宁中医药大学学报,2006,8(5):5-6.
16.秦银忠,黄晓峰,李志刚.清藏固本汤配合化疗治疗中晚期大肠癌疗效观察[J].现代中西医结合杂志,2006,15(24):3377-3378.
17.赵文硕,张青,等.加味升血汤配合化疗治疗中晚期大肠癌的临床研究[J].国际中医中药杂志,2006,28(5):306-308.
18.蒋益兰,潘博,仇湘中.健脾消癌饮配合化疗治疗大肠癌术后40例总结[J].湖南中医杂志,2001,17(5):9-10.
19.王振飞.大肠癌术后化疗配合艾迪注射液临床观察[J].福建中医药,2007,38(4):9-10.
20.尤建良,周留勇.扶正和胃合剂治疗大肠癌48例[J].药学与临床研究 2007,15(1):60-62.
21.杨彦,刘碧清,杜秋霞.肠复康胶囊治疗中晚期大肠癌55例临床观察[J].江苏中医药,2005,26(1):18-20.
22.金哲秀.针灸两步法治疗大肠癌27例临床分析.上海中医药杂志2003,37:48-49.
23.穆立新,姜军作,范杰华.针灸治疗大肠癌术后30例疗效观察.大连大学学报,2006,27:100-106.
24.黄兆明.中药灌肠综合治疗癌性肠梗阻临床观察[J].实用中西医结合临床,2003,3(3):36.
25.郑玉玲,王新杰.肠达顺灌肠液治疗湿热蕴结型大肠癌的临床研究[J].河南中医,2002,22(1):12.