不同种类生长抑素类似物对肝细胞肝癌作用差异的实验研究

吕修和; 王春晖; 谢艳

不同种类生长抑素类似物对肝细胞肝癌作用差异的实验研究

Differences of Therapeutic Efficacy Between Different Kinds of Somatostatin Analogue for Primary 

摘要

摘要: 目的观察SOM230、奥曲肽、兰瑞肽对体内外Bel-7402肝癌细胞的作用，分析其疗效差异及可能的机制。方法采用不同浓度（1×10-10～1×10-5mol/L）SOM230、奥曲肽和兰瑞肽处理Bel-7402肝癌细胞，作用24 h、48 h和72 h时，采用CCK8法检测细胞增殖；采用1×10-5mol/L 浓度的3种药物处理细胞24 h、48 h和72 h， Annexin Ⅴ-FITC/PI双染法检测细胞凋亡；荧光定量PCR检测药物作用前后细胞生长抑素受体（SSTR）表达的变化。建立Bel-7402细胞裸鼠移植瘤模型，干预组分别予SOM230、奥曲肽、兰瑞肽100 μg/（kg·d），对照组予同体积生理盐水，连续用药6周（皮下注射），荧光定量PCR及Western blot检测肿瘤组织中SSTR的基因及蛋白表达变化。结果3种药物均可抑制Bel-7402细胞的增殖，但均无法促进其凋亡。在体外水平，SSTR1、SSTR4在各组间的表达量均不随时间发生变化；与对照组比较，SSTR2在干预组中的表达量均表现为随干预时间延长而减少(PSSTR5在干预组中的表达量均表现为随干预时间延长先增加后减少(PSSTR1基因在SOM230组表达量较奥曲肽、兰瑞肽及对照组增加；SSTR2基因在3个干预组中的表达量较对照组均增加；SSTR5基因在SOM230组及兰瑞肽组中的表达量较奥曲肽组及对照组增加；上述差异均具统计学意义(P<0.05)。Western blot从蛋白水平验证了上述结果。结论在一定浓度范围内，采用针对SSTR2和SSTR5具有高亲合力的生长抑素类似物（SSTA）长期治疗可抑制肝癌细胞的生长。

Abstract: Objective To explore the effects of SOM230, octreotide and lanreotide on hepatocellular carcinoma cell line Bel-7402 In vivoand In vitro, and to analyze the differences of their therapeutic efficacy with relevant mechanisms. Methods At different time points (24, 48, 72 h), the cell counting kit-8 (CCK8) was used to evaluate cell proliferation (drug concentration 1×10-10-1×10-5mol/L) and the Annexin Ⅴ-FITC/PI staining was used to assess cell apoptosis (drug concentration 1×10-5mol/L), while the real-time quantitative PCR was used to detect cellular SSTRexpression changes before and after interventions. A transplanted tumor model was set up, and the tumor-bearing nude mice were treated by three drugs with a common dose of 100 μg/（kg·d） or same volume of normal saline, respectively. After a treatment period of 6 weeks, real-time quantitative PCR and Western blot test were performed to detect the expression changes ofSSTR in tumor tissues from the level of gene and protein. Results All three drugs could inhibit the cell proliferation of Bel-7402. However, they were unable to promote the cell apoptosis. In vitro, the expressions ofSSTR1, SSTR4genes did not change over time in each group. The expressions ofSSTR2gene were decreased in three intervention groups while the expressions ofSSTR5gene were increased first and then decreased. Compared with the control group, all differences have statistical significance (PIn vivo, the expression ofSSTR1gene in SOM230 group was increased when compared with that of the other groups; the expressions ofSSTR2gene in three intervention groups were increased when compared with that of the control group; the expressions of SSTR5gene in SOM230 group and lanreotide group were increased when compared with that of the octreotide group and the control group, and all differences have statistical significance (P<0.05). The Western blot test confirmed these results from the protein level. Conclusion In a certain concentration range, the long-term treatment of SSTA with high affinities to SSTR2 and SSTR5 could inhibit the growth of HCC.

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