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450003 郑州,郑州大学人民医院消化内科
[韩双印,E-mail:hansyzzu@163.com HAN Shuangyin, E-mail: hansyzzu@163.com
收稿:2017-12-01,
修回:2018-01-12,
纸质出版:2018-07-15
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张立达, 房新辉, 杨玉秀, 等. 盐酸千金藤碱抑制肝癌肾脏、睾丸转移及其分子机制[J]. 第三军医大学学报, 2018,40(13):1192-1197.
Lida ZHANG, Xinhui FANG, Yuxiu YANG, et al. Cepharanthine hydrochloride inhibits kidney and testicular metastases from hepatocellular carcinoma in xenograft tumor models in nude mice[J]. Journal of Third Military Medical University, 2018, 40(13): 1192-1197.
张立达, 房新辉, 杨玉秀, 等. 盐酸千金藤碱抑制肝癌肾脏、睾丸转移及其分子机制[J]. 第三军医大学学报, 2018,40(13):1192-1197. DOI: 10.16016/j.1000-5404.201712207.
Lida ZHANG, Xinhui FANG, Yuxiu YANG, et al. Cepharanthine hydrochloride inhibits kidney and testicular metastases from hepatocellular carcinoma in xenograft tumor models in nude mice[J]. Journal of Third Military Medical University, 2018, 40(13): 1192-1197. DOI: 10.16016/j.1000-5404.201712207.
目的
2
通过裸鼠肝原位移植荧光人肝癌细胞,观察肝癌细胞肾脏、睾丸转移情况,并探讨盐酸千金藤碱抑制肝癌细胞转移及其分子机制。
方法
2
采用慢病毒转染构建稳定表达荧光素酶基因的人肝癌细胞SMCC-7721-Luc。BALB/c裸鼠皮下接种SMCC-7721-Luc细胞,待肿瘤长至100 mm
3
左右,取皮下瘤植入裸鼠肝脏包膜,建立裸鼠原位肝癌模型。裸鼠腹腔注射盐酸千金藤碱,用小动物活体成像系统观察肝原位肿瘤肾脏、睾丸转移情况。给药21 d后,解剖取肿瘤组织,Western blot检测相关蛋白表达,同时取肝脏、肾脏、睾丸等组织,用10%中性甲醛固定,进行病理分析。
结果
2
构建的SMCC-7721-Luc细胞能稳定表达Luc基因,裸鼠肝原位接种该细胞后产生较强、稳定的荧光。活体成像系统显示盐酸千金藤碱能抑制裸鼠原位肝癌向肾脏、睾丸转移,与其降低MMP-7和N-cadherin蛋白表达,上调E-cadherin蛋白表达有关。
结论
2
盐酸千金藤碱通过调控MMP-7、N-cadherin和E-cadherin蛋白抑制原位肝癌向肾脏、睾丸转移。
Objective
2
To observe the kidney and testicular metastases from hepatocellular carcinoma in nude mouse model by orthotopic transplantation of human liver cancer cells stably expressing reporter gene luciferase
and investigate the anti-metastatic effects of cepharanthine hydrochloride (CEH) and its molecular mechanism.
Methods
2
SMMC-7721 cells were transfected with luciferas
e gene mediated by lentiviral vector to prepare SMMC-7721-Luc cells. Then the obtained cells were inoculated subcutaneously into the back of BALB/c nude mice to establish xenograft tumor model of nude mice. Till the masses grow into a size of 100 mm
3
the masses were collected and cut into blocks of 2 mm×1 mm×1 mm
and then transplanted into the liver to establish orthotopic tumor model of liver cancer. The tumor growth and kidney and testicular metastases were observed in nude mice after intraperitoneal injection of CEH by bioluminescence imaging system. In 21 d after CEH injection
the tumor tissues were harvested for expression of related proteins by Western blotting
and the liver
kidneys and testis were collected and fixed with 10% neutral formaldehyde for pathological observation.
Results
2
The established SMMC-7721-Luc cells stably expressed luciferase gene
and produced strong and stable fluorescence after orthotopic transplantation in nude mice. The bioluminescence imaging system displayed that CEH significantly inhibited the kidney and testicular metastases of orthotopic liver tumor
which was associated with the down-regulation of MMP-7 and N-cadherin
and up-regulation of E-cadherin.
Conclusion
2
CEH significantly inhibits the kidney and testicular metastases from hepatocellular carcinoma by regulating MMP-7
N-cadherin and E-cadherin.
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