Effect Of Knocking Down The ECEL1 Gene On The Growth Of Hepatocellular Carcinoma Cells In Nude Mice

Objective:To investigate the effect of knocking down the ECEL1 gene on the growth of hepatocellular carcinoma cells(BEL-7404)in vivo,and to provides new evidence for targeted treatment of hepatocellular carcinoma in clinic.Methods:(1)Preparation of tumor-forming cells: The hepatocellular carcinoma cell line(BEL-7404)was cultured and passaged,and human hepatocellular carcinoma cells in the logarithmic growth phase were used for experiments.Use CON053 lentivirus and LV-ECEL1-RNAi(48784-1)lentivirus infected hepatocellular carcinoma cell line(BEL-7404),and a blank control group was set up.The infection efficiency was observed under a fluorescence microscope 72 hours later.(2)Animal tumor formation experiment:Twenty nude mice were numbered and grouped: Nude mice of control group were injected with human hepatocellular carcinoma cells without knockdown of ECEL1.Nude mice of experimental group,were injected with human hepatocellular carcinoma cells that had knocked down the ECEL1.(3)Observe the changes of the subcutaneous tumors in the twogroups of nude mice,record the tumor length and diameter,tumor weight and calculate the tumor volume data for comparison and analysisResults:(1)Results of preparation of tumor-forming cells:72 hours after infection,no fluorescence expression was observed in the blank control group under a fluorescence microscope;Human hepatocellular carcinoma cells(BEL-7404)infected with the ECEL1 interference plasmid lentivirus(KD group)and human hepatocellular carcinoma cells(BEL-7404)(NC group)infected with the negative control virus CON053 are clearly visible Green fluorescence.The fluorescence rate of both groups was above 80%,and the cell status was normal.This indicates that the tumor cells were successfully prepared.(2)Changes in tumor volume of subcutaneously transplanted hepatocellular carcinoma in mice:Twenty-five days after subcutaneous injection of human hepatocellular carcinoma cells(BEL-7404),tumors formed in 10 nude mice in the control group,with a tumor formation rate of 100%,and tumors formed in 10 nude mice in the experimental group,with a tumor formation rate.100%.During the observation period,the tumor volume of the nude mice in the two groups kept increasing with time.Comparing the tumor volume at 5 time points,it was concluded that the experimental group(KD group)increased more slowly than the control group(NC group),and the tumor volume at different time points in the two groups were significantly different(p<0.05).(3)Changes of fluorescenceexpression in subcutaneously transplanted hepatocellular carcinoma in mice:Experimental group(KD group)the total fluorescence expression in the tumor area(2.50E+10)were significantly reduced compared with the control group(2.82E+10),and the difference was significant(p=4.8337E-06).(4)Changes in tumor weight of subcutaneously transplanted hepatocellular carcinoma in mice: After euthanizing the mice,the tumor tumors were removed and weighed.The results showed that the tumor weight of the KD group(0.457±0.220)was significantly reduced compared with the NC group(0.780 ± 0.204)(p <0.05).Conclusion: Knockdown of the ECEL1 gene can inhibit the growth rate of human hepatocellular carcinoma cells(BEL-7404)in nude mice.