肝细胞癌射频消融体外热疗残癌细胞模型c-Met蛋白的表达变化
发布时间：2019-05-05 17:18
【摘要】：目的:c-Met蛋白与肿瘤细胞的生长、增殖、迁移、侵袭及凋亡的保护等密切相关。本实验研究通过建立体外热疗残癌细胞HepG2模型,模拟肝细胞癌(hepatocellular carcinoma,HCC)经射频消融(radiofrequency ablation,RFA)治疗后残留癌细胞,利用免疫细胞化学方法检测热疗后残留癌细胞中c-Met蛋白的表达变化,旨在探讨高温热疗对残癌细胞c-Met蛋白表达的影响,明确c-Met蛋白对残癌细胞的作用,为RFA后HCC复发转移的机制和以c-Met为目标靶点的药物研究提供理论依据与实验基础。方法:1、将HePG2细胞放入正常浓度210 mL/L氧气、50g/L二氧化碳、饱和湿度以及37°C的培养箱中培养。培养液为含10%胎牛血清的DMEM细胞培养基,隔日更换培养液。人肝癌细胞HepG2呈贴壁生长。实验所用细胞均处于对数生长期。2、实验组体外热疗残余癌细胞模型的建立:将对数生长期的HePG2细胞置于含正常浓度210 mL/L氧气、50g/L二氧化碳及饱和湿度的细胞培养箱内。培养液为含10%胎牛血清的DMEM细胞培养基。通过调整培养箱的温度使其分别达到41℃、45℃、50℃,根据培养温度的不同,再依次分为A、B、C三个组,各于6 hs、12hs、24 hs、48 hs观察HePG2细胞生长情况,残留癌细胞进行后续实验。3、对照组残癌细胞模型的建立:将对数生长期的HePG2细胞置于含正常浓度210 mL/L氧气、50g/L二氧化碳及饱和湿度的细胞培养箱内。培养液为含10%胎牛血清的dmem细胞培养基。保持培养箱的温度为37℃,于6hs、12hs、24hs、48hs观察hepg2细胞生长情况,残留癌细胞进行后续实验。4、免疫细胞化学方法检测各组hepg2残癌细胞c-met蛋白的表达。5、spss软件统计学分析:(1)采用χ2检验比较实验组a、b、c两两间c-met蛋白表达阳性率的差异;(2)采用χ2检验比较各实验组与对照组间c-met蛋白表达阳性率的差异。(p0.05)结果:1、经免疫细胞化学方法检测,在光镜下c-met蛋白阳性细胞为hepg2胞膜呈棕黄色染色,而胞核不染色。2、c-met蛋白在体外热疗残癌细胞模型实验组a、b、c中表达的阳性率依次为33.2%、35.7%、36.5%。虽然三者阳性率都升高,但a与b、b与c、a与c之间依次比较,差异在统计学上都没有意义(pab=0.1440.05,pbc=0.6480.05,pac=0.0550.05)。说明当热疗温度高于某一值(如37℃)之后,c-met蛋白阳性表达率并不一定随温度升高而升高,与热疗温度之间无依赖性。3、c-met蛋白在体外热疗残癌细胞模型实验组a、b、c中表达的阳性率依次为33.2%、35.7%、36.5%,对照组中c-met蛋白表达的阳性率为10.5%。a与对照组比较差异有统计学意义(pa0.001),b与对照组比较差异有统计学意义(pb0.001),c与对照组比较差异有统计学意义(pc0.001)。与对照组相比,实验组中残癌细胞c-met蛋白表达的阳性率更高,说明热疗能促进残癌细胞c-met蛋白的表达。结论:1、体外热疗残癌细胞hepg2中c-met蛋白表达的阳性率更高,高温热疗可以促进残癌细胞c-met蛋白的表达。2、c-met蛋白可能成为提示肝细胞癌复发、转移及疗效的一个新的生物标志物。
[Abstract]:Aim: c-Met protein is closely related to the growth, proliferation, migration, invasion and apoptosis protection of tumor cells. In this study, the residual cancer cells HepG2 model in vitro hyperthermia was established to simulate the residual cancer cells after radiofrequency ablation (radiofrequency ablation,RFA) in hepatocellular carcinoma (hepatocellular carcinoma,HCC). The expression of c-Met protein in residual cancer cells after hyperthermia was detected by immunocytochemistry. The purpose of this study was to investigate the effect of hyperthermia on the expression of c-Met protein in residual cancer cells and to clarify the effect of c-Met protein on residual cancer cells. It provides a theoretical basis and experimental basis for the mechanism of recurrence and metastasis of HCC after RFA and the study of drugs targeting c-Met. Methods: 1. HePG2 cells were cultured in a incubator with normal concentration of 210 mL/L oxygen, 50g/L carbon dioxide, saturated humidity and 37 掳C. DMEM cell culture medium containing 10% fetal bovine serum was used in the culture medium, and the culture medium was changed every other day. Human hepatocellular carcinoma cell line HepG2 grew adherently. The cells used in the experiment were in the logarithmic growth phase. 2. In vitro hyperthermia model of residual cancer cells in the experimental group was established: the logarithmic growth phase of HePG2 cells was placed in the normal concentration of 210mL/L oxygen. 50g/L carbon dioxide and saturated humidity in the cell culture box. The culture medium was DMEM cell medium containing 10% fetal bovine serum. By adjusting the temperature of the incubator to 41 鈩，
本文编号：2469765
[Abstract]:Aim: c-Met protein is closely related to the growth, proliferation, migration, invasion and apoptosis protection of tumor cells. In this study, the residual cancer cells HepG2 model in vitro hyperthermia was established to simulate the residual cancer cells after radiofrequency ablation (radiofrequency ablation,RFA) in hepatocellular carcinoma (hepatocellular carcinoma,HCC). The expression of c-Met protein in residual cancer cells after hyperthermia was detected by immunocytochemistry. The purpose of this study was to investigate the effect of hyperthermia on the expression of c-Met protein in residual cancer cells and to clarify the effect of c-Met protein on residual cancer cells. It provides a theoretical basis and experimental basis for the mechanism of recurrence and metastasis of HCC after RFA and the study of drugs targeting c-Met. Methods: 1. HePG2 cells were cultured in a incubator with normal concentration of 210 mL/L oxygen, 50g/L carbon dioxide, saturated humidity and 37 掳C. DMEM cell culture medium containing 10% fetal bovine serum was used in the culture medium, and the culture medium was changed every other day. Human hepatocellular carcinoma cell line HepG2 grew adherently. The cells used in the experiment were in the logarithmic growth phase. 2. In vitro hyperthermia model of residual cancer cells in the experimental group was established: the logarithmic growth phase of HePG2 cells was placed in the normal concentration of 210mL/L oxygen. 50g/L carbon dioxide and saturated humidity in the cell culture box. The culture medium was DMEM cell medium containing 10% fetal bovine serum. By adjusting the temperature of the incubator to 41 鈩，
本文编号：2469765
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2469765.html
