蟾毒灵对人膀胱癌T24细胞增殖及细胞凋亡的影响
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摘要
目的:探讨蟾毒灵对人膀胱癌T24细胞在细胞增殖、细胞周期、细胞凋亡等方面的影响及其可能的机制,为膀胱癌的治疗寻找新的策略。方法:将实验分为空白组和蟾毒灵组(蟾毒灵组分为不同浓度组)。用CCK-8法评价蟾毒灵对人膀胱癌细胞株T24细胞体外增殖的影响,流式细胞术(flow cytometry, FCM)测定各组细胞周期及凋亡率的变化。Western-blot法检测凋亡相关蛋白Caspase-3、Caspase-9、Bax及Bcl-2的表达。裸鼠成瘤实验检测蟾毒灵对人膀胱癌细胞株T24细胞体内生长的影响。结果:与空白组比较,蟾毒灵可抑制膀胱癌T24细胞生长,浓度、时间依赖性明显,并将细胞周期阻滞于G0/G1期。蟾毒灵有明显诱导膀胱癌T24细胞凋亡作用,其凋亡作用随药物浓度增加而增加。Western-blot结果显示:蟾毒灵可明显上调Caspase-3、Caspase-9、Bax蛋白的表达,下调Bcl-2蛋白的表达。裸鼠皮下瘤生长实验示:蟾毒灵不同浓度干预组细胞种植瘤的平均体积均明显低于空白组,且蟾毒灵浓度越高,其抑制瘤体生长的趋势越明显。结论:蟾毒灵可抑制人膀胱癌T24细胞增殖,并将细胞周期阻滞在G0/G1期,可显著诱导T24细胞凋亡,并能有效抑制裸鼠模型肿瘤的生长。其作用机制可能是上调凋亡蛋白Caspase-3、Casepase-9、Bax的表达并下调凋亡抑制蛋白Bcl-2的表达等。
Objective: To explore the effect of bufalin on cell proliferation, cell cycle and apoptosis of human bladder cancer cell line T24 and its possible mechanism, so as to find a new strategy for the treatment of bladder cancer. Methods: Nude mice were divided into blank control group and Bufalin poison group(Bufalin poison group was divided into different concentration groups). CCK-8 was used to observe the inhibition of bufalin impact on human bladder cancer cell line T24 cells grown in vitro, FCM was used to detect cell cycle profile and apoptosis rate of T24, Western-blot method to detect the expression of apoptosis-related proteins caspase-3, caspase-9, Bax and Bcl-2. The effect of bufalin on the growth of human bladder cancer cell line T24 in vivo was examined by nude mice tumorigenesis experiment. Results: The growth of T24 cells was inhibited by bufalin in a dose-and time-dependent manner, which may provide the cell cycle arrest at the G0/G1 phase. Compared with the control group, bufalin could significantly induced apoptosis. Western-blot results showed that: Bufalin can up-regulate the expression of caspase-3, caspase-9 and Bax, and down-regulated the expression of Bcl-2 protein. Subcutaneous tumor growth experiment showed that the average volume of cell implant tumor was significantly lower than that of the blank group in different concentration intervention group,and the higher the concentration of bufalin, the more obvious its tendency to inhibit the growth of tumor body.Conclusion: Bufalin can inhibit the proliferation of human bladder cancer T24 cells, and arrest the cells in G0/G1 phase. Moreover, bufalin can significantly induce apoptosis in T24 cells. And it can effectively inhibit the growth of nude mouse model tumor. Its mechanism of induction of apoptosis may be upregulated caspase-3,caspase-9, Bax protein expression and down-regulates the expression of Bcl-2 protein.
Objective: To explore the effect of bufalin on cell proliferation, cell cycle and apoptosis of human bladder cancer cell line T24 and its possible mechanism, so as to find a new strategy for the treatment of bladder cancer. Methods: Nude mice were divided into blank control group and Bufalin poison group(Bufalin poison group was divided into different concentration groups). CCK-8 was used to observe the inhibition of bufalin impact on human bladder cancer cell line T24 cells grown in vitro, FCM was used to detect cell cycle profile and apoptosis rate of T24, Western-blot method to detect the expression of apoptosis-related proteins caspase-3, caspase-9, Bax and Bcl-2. The effect of bufalin on the growth of human bladder cancer cell line T24 in vivo was examined by nude mice tumorigenesis experiment. Results: The growth of T24 cells was inhibited by bufalin in a dose-and time-dependent manner, which may provide the cell cycle arrest at the G0/G1 phase. Compared with the control group, bufalin could significantly induced apoptosis. Western-blot results showed that: Bufalin can up-regulate the expression of caspase-3, caspase-9 and Bax, and down-regulated the expression of Bcl-2 protein. Subcutaneous tumor growth experiment showed that the average volume of cell implant tumor was significantly lower than that of the blank group in different concentration intervention group,and the higher the concentration of bufalin, the more obvious its tendency to inhibit the growth of tumor body.Conclusion: Bufalin can inhibit the proliferation of human bladder cancer T24 cells, and arrest the cells in G0/G1 phase. Moreover, bufalin can significantly induce apoptosis in T24 cells. And it can effectively inhibit the growth of nude mouse model tumor. Its mechanism of induction of apoptosis may be upregulated caspase-3,caspase-9, Bax protein expression and down-regulates the expression of Bcl-2 protein.
引文
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[5]林文彬.膀胱癌治疗方法的选择[J].当代医学,2012,18(8):101-103.
[6]赵国平,戴慎,陈仁涛,等.中药大辞典(下册)[M].2版.上海:上海科学技术出版社,2006:3832-3834.
[7] Yu C H,Kan S F,Pu H F, et al. Apoptotic signaling in bufalin and cinobufagin treated androgen-dependent and independant human prostate cancer cells[J].Cancer Sci,2008,99(12):2467-2476.
[8] Fanghua Qi,Yoshinori Inagaki,Bo Gao,et al. Bufalin and cinobufagin induce apoptosis of human hepatocellular carcinoma cells via Fas-and mitochondria-mediated pathways[J].Cancer Sci,2011,102(5):951-958.
[2] Ding Da Wei,Zhang Yong Hong,Huang Xin En,et al.Bufalin Induces Mitochondrial Pathway-Mediated Apoptosis in Lung Adenocarcinoma Cells[J].Asian Pacific Journal of Cancer Prevention,2014,15(23):10495-10500.
[3] Jemal A, Bray F, Center M M, et al. Global cancer statistics[J].CA Caneer J Clin,2011,61(2):69-90.
[4]韩苏军,张思维,陈万青,等.中国膀胱癌死亡现状及流行趋势分析[J].现代泌尿外科杂志,2013,18(3):228-232.
[5]林文彬.膀胱癌治疗方法的选择[J].当代医学,2012,18(8):101-103.
[6]赵国平,戴慎,陈仁涛,等.中药大辞典(下册)[M].2版.上海:上海科学技术出版社,2006:3832-3834.
[7] Yu C H,Kan S F,Pu H F, et al. Apoptotic signaling in bufalin and cinobufagin treated androgen-dependent and independant human prostate cancer cells[J].Cancer Sci,2008,99(12):2467-2476.
[8] Fanghua Qi,Yoshinori Inagaki,Bo Gao,et al. Bufalin and cinobufagin induce apoptosis of human hepatocellular carcinoma cells via Fas-and mitochondria-mediated pathways[J].Cancer Sci,2011,102(5):951-958.