蝎毒粗提物对人乳腺癌细胞MCF-7的抑制作用及机制
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摘要
恶性肿瘤是严重威胁人类生命安全的疾病之一。寻求高效低毒的抗肿瘤药物及治疗方法一直是科学界研究的热点。蝎毒作为一种生物毒素,凭借其广泛的生理学效应,极具研究价值。本文旨在研究蝎毒对肿瘤细胞的杀伤作用和敏感细胞系的筛选,并对其作用机制进行探讨,为蝎毒作为抗肿瘤药物的开发及临床应用提供实验依据。
目的:观察蝎毒粗提物对人乳腺癌细胞MCF-7的抑制作用,初步探讨其作用机制。
方法:采用MTT法和荧光免疫技术在三种常见肿瘤细胞(人肝癌细胞SMMC-7721、人乳腺癌细胞MCF-7和人宫颈癌细胞Hela)中选出对蝎毒敏感性较高的肿瘤细胞。免疫细胞化学法检测药物干预后,两种相关凋亡蛋白Caspase-3和Bcl-2在蛋白水平的表达量。流式细胞仪观察药物对细胞周期的影响,Western Blotting检测药物干预后,细胞周期相关蛋白CyclinD1在蛋白水平表达的变化。
结果:MTT结果显示600μg/ml蝎毒粗提物对7721和MCF-7细胞均有抑制作用,与阴性对照组相比p<0.05.免疫荧光技术显示7721细胞对蝎毒的敏感性好于Hela细胞。免疫细胞化学检测显示药物干预后,MCF-7细胞中Caspase-3表达上调,Bcl-2表达下调。流式细胞仪显示药物干预后,处于S期细胞数减少,G0/G1期细胞数增多,与阴性对照组相比p<0.05。Western Blotting显示细胞周期相关蛋白CyclinD1蛋白表达量减少。
结论:蝎毒粗提物可抑制MCF-7细胞的生长和增殖,其作用机制可能与诱导细胞发生凋亡,影响细胞G0/G1期和S期有关,在乳腺癌治疗中具有重要价值。
Cancer is a serious threat to human life and safety of one of the diseases. To seek efficient and low toxicity of anticancer drμgs and treatment of the scientific community has been on the hot spots. Scorpion venom is a biological toxin, because of its wide range of physiological effects closely investigated. This paper aims to study the destruction about scorpion venom on human breast cancer cell line MCF-7 and sensitive cell line, and their mechanisms were discussed for the development of scorpion toxin of anti-tumor clinical applications experimental basis.
Objective: detected growth inhibition of using the extractive of scorpion venom on human breast cancer cell line MCF-7, and investigated the mechanism.
Methods: bolting the sensitive cell line in hepatoma cell SMMC-7721、Hela cell and breast cancer cell MCF-7 by MTT and IMF; detecting the apoptosis-related proteins expressed levels of caspase-3 and Bcl-2 proteins with ICC; observed the effect on cell cycle by FCM; detecting the cell cycle-related protein expressed levels of cyclinD1 with western blotting. Results: MTT method displayed that the inhibition on 7721 and MCF-7 by using the 600μg/ml extractive of scorpion venom, compared with the negative group(p<0.05);IMF method displayed that the sensitive to scorpion venom of 7721 cell was better than Hela cell after drug intervention; ICC method displayed that apoptosis protein caspase-3 increased and anti-apoptosis protein Bcl-2 decreased after drug intervention; FCM displayed that the amount of S period of time decreased, the amount of G1 period time increased after drug intervention, compared with the negative group(p<0.05); western blotting displayed that the expression of cell cycle-related protein cyclinD1 decreased after drug intervention.
Conclusion: the extractive of scorpion venom inhibited the growth of human breast cancer MCF-7, and induced apoptosis in MCF-7 cell, the mechanism maybe related to G1 and S period of time, there was a significant value in treating human breast cancer.
目的:观察蝎毒粗提物对人乳腺癌细胞MCF-7的抑制作用,初步探讨其作用机制。
方法:采用MTT法和荧光免疫技术在三种常见肿瘤细胞(人肝癌细胞SMMC-7721、人乳腺癌细胞MCF-7和人宫颈癌细胞Hela)中选出对蝎毒敏感性较高的肿瘤细胞。免疫细胞化学法检测药物干预后,两种相关凋亡蛋白Caspase-3和Bcl-2在蛋白水平的表达量。流式细胞仪观察药物对细胞周期的影响,Western Blotting检测药物干预后,细胞周期相关蛋白CyclinD1在蛋白水平表达的变化。
结果:MTT结果显示600μg/ml蝎毒粗提物对7721和MCF-7细胞均有抑制作用,与阴性对照组相比p<0.05.免疫荧光技术显示7721细胞对蝎毒的敏感性好于Hela细胞。免疫细胞化学检测显示药物干预后,MCF-7细胞中Caspase-3表达上调,Bcl-2表达下调。流式细胞仪显示药物干预后,处于S期细胞数减少,G0/G1期细胞数增多,与阴性对照组相比p<0.05。Western Blotting显示细胞周期相关蛋白CyclinD1蛋白表达量减少。
结论:蝎毒粗提物可抑制MCF-7细胞的生长和增殖,其作用机制可能与诱导细胞发生凋亡,影响细胞G0/G1期和S期有关,在乳腺癌治疗中具有重要价值。
Cancer is a serious threat to human life and safety of one of the diseases. To seek efficient and low toxicity of anticancer drμgs and treatment of the scientific community has been on the hot spots. Scorpion venom is a biological toxin, because of its wide range of physiological effects closely investigated. This paper aims to study the destruction about scorpion venom on human breast cancer cell line MCF-7 and sensitive cell line, and their mechanisms were discussed for the development of scorpion toxin of anti-tumor clinical applications experimental basis.
Objective: detected growth inhibition of using the extractive of scorpion venom on human breast cancer cell line MCF-7, and investigated the mechanism.
Methods: bolting the sensitive cell line in hepatoma cell SMMC-7721、Hela cell and breast cancer cell MCF-7 by MTT and IMF; detecting the apoptosis-related proteins expressed levels of caspase-3 and Bcl-2 proteins with ICC; observed the effect on cell cycle by FCM; detecting the cell cycle-related protein expressed levels of cyclinD1 with western blotting. Results: MTT method displayed that the inhibition on 7721 and MCF-7 by using the 600μg/ml extractive of scorpion venom, compared with the negative group(p<0.05);IMF method displayed that the sensitive to scorpion venom of 7721 cell was better than Hela cell after drug intervention; ICC method displayed that apoptosis protein caspase-3 increased and anti-apoptosis protein Bcl-2 decreased after drug intervention; FCM displayed that the amount of S period of time decreased, the amount of G1 period time increased after drug intervention, compared with the negative group(p<0.05); western blotting displayed that the expression of cell cycle-related protein cyclinD1 decreased after drug intervention.
Conclusion: the extractive of scorpion venom inhibited the growth of human breast cancer MCF-7, and induced apoptosis in MCF-7 cell, the mechanism maybe related to G1 and S period of time, there was a significant value in treating human breast cancer.
引文
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2.王春光,戚正武,等.蝎长链神经毒秦研究进展.生命科学研究2001 ,5(2):95-101
3.余茂耘,韦传宝,等蝎毒的生理活性成分及临床应用.中国临床康复2004,8(9):1754-1755
4.苏智广,梁爱华.东亚钳蝎神经毒素的研究进展.山西大学学报1997 ,20(3):343-347.
5.汤耀光,陈斌,梁业玲,等.蛇毒多肽和蝎毒多肽对小鼠肝癌移植瘤的抑制作用和Caspase-3表达的影响.广东医学院学报2006,34(5):18-21
6.宋红霞,杨建斌,等.蝎毒抗癌多肽(APBMV)与5-Fu序贯用药治疗小鼠肝癌.长治医学院学报2006,20(1)
7.韩雪飞,王永奎,陈华艳,等.蝎毒抗癌多肽纯化组分Ⅲ对小鼠肝癌的生长抑制作用及带瘤小鼠胸腺重量的影响.河南医科大学学报2000;35(4)
8.李建伟,胡静,张桂荣,等.东亚钳蝎蝎毒多肽组分Ⅲ对肝癌细胞凋亡的影响.吉林大学学报2006,32(4):625-628
9.李桂生,杨春旭,朱灵,等.蝎毒对人肝癌Bel-7404细胞端粒酶活性的影响.东南大学学报2004,23(3)
10.魏玲,董伟华,孔天翰, ,等.东亚钳蝎毒抗癌多肽对人肿瘤细胞系和动物移植瘤的作用.中国癌症杂志2002,12(2)
11.杨春旭,韦小谋,等.蝎毒与丝裂霉素联合应用对人肝癌细胞生长的影响.东南大学学报2005,24(4)
12.杨春旭,戴盛明,朱灵,等.东亚钳蝎毒生物提取物对人肝癌细胞体外增殖及凋亡的影响.东南大学学报2002,19(2)
13.解霞,赵瑾瑶,高清波,等.蝎毒对人乳腺癌MCF-7/ADM细胞多药耐药的影响机机制研究.大连医科大学学报2006,28(5)
14.于丽君,孔力.蝎毒逆转人乳腺癌细胞株耐药性研究.吉林大学学报2004;30(2)
15.付士波,杨英,闫凤琴,等.蝎毒组分Ⅰ对卵巢癌细胞SKOV3生长的抑制作用.吉林大学学报2007;33(1):41-43
16.王兆朋,张维东,张捷,等.蝎毒多肽提取物对非激素依赖性前列腺癌细胞增殖抑制作用的实验研究.中国药理学通报2006;22(8)
17.张月英,张维东,贾青,等.蝎毒多肽提取物诱导前列腺癌DU-145细胞凋亡的实验研究.实用癌症杂志2006;21(3):225-226
18.赵灿国,董伟华,等.蝎毒及其分离组分对4种肿瘤细胞生长的抑制作用.广州医学院学报2006;34(1)
19.王秀丽,李冬晖,等.东亚钳蝎毒抗癌多肽抗肿瘤作用的体内试验.实用医药杂志2003;20(2)
20.贾金秋,王颖,张维东,等.蝎毒多肽提取物对肺癌荷瘤裸鼠血流变学的影响.山东医药2006;46(27):11-12
21.辜为高,邱庐山,等.蝎毒多肽Ⅲ和蛇毒多肽Ⅷ对小鼠Lewis肺癌生长和肺部转移的抑制作用.广州医学院学报2005;33(6)
22.沈东海,高春芳,王元和,等.蝎毒对人大肠癌细胞株体外抑制杀伤作用的观察.实用癌症杂志2002;17(4)
23.孔天翰,林山.蝎毒多肽对肿瘤细胞的抑制作用研究.中国病理生理杂志2004,20(6):968～972
24.贾莉,孟秀香,等.蝎毒对白血病HL-60细胞生长的抑制和诱导凋亡作用研究.中药药理与临床2001;17(6)
25.贾莉,孙宏丹,孟秀香,等.蝎毒对白血病Raji细胞株的生长抑制和诱导凋亡作用的研究.白血病·淋巴瘤2001:10(4)
26.吴孔明,李醒亚,李安平,等.蝎毒抗癌因子外用治疗癌性溃疡6例.中国中西医结合杂志1998;(8)
1.周新华.生物化学与生物物理进展1984,11(2)20
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