Effects Of RNA Interference LI-cadherin On The Biological Behavior Of Hep3B Hepatoma Cell Line In Vitro

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors, which has the high grade malignancy, progress rapidly, metastasis early, grow infiltratively, also has the unfavorable prognosis. The resection is the main therapy method, but the prognosis is very poor. The abnormal adhesion expression of the tumour cell is the key factor of the tumor metastasis and recurrence. To study the adhesion molecule is the only reliable way to learn the infiltration and the metastasis of the HCC.Cadherin is a calcium-dependent adhesion glycoprotein family mediating the adhesion between cells and homologous cells, studies have shown that cadherin plays an important role in the stretching and moving, signal transduction and activation, growth and differentiation of the cells, wound healing and tumor metastasis and other physiological and pathological activities. So the metastasis and invasiveness of the HCC are largely related to the defects of cadherin's structure and function.LI-cadherin-SiRNA was transfected to hepatumor cells Hep3B in this experiment. To investigate the effects on the ability of growth, adhesion and metastasis of Hep3B hepatumor cell line after the expression of LI-cadherin was inhibited. Objective:To observe the changes of growth, adhesion and metastasis of Hep3B after LI-cadherin SiRNA was transfected.Methods:We transfect LI-cadherin siRNA to Hep3B, then observe the-ransfection efficiency. Detect the changes of LI-cadherin gene and protein by RT-PCR and Western blot during the process; examine the effect of LI-cadherin on the growth and adhesion of HCC by MTT, and the metastasis of HCC by Boyden cell invasion test.Results:1. Green fluorescence could be seen from the hepatocellular cells Hep3B that transfected with vector LI-cadherin-SiRNA, which proved that the transfection is well. The proportion was calculated of fluorescence cells of all the cells, which was 80%, which complyed with requirements of experiments.2. Real-time PCR and Western blot were used to detect the expression of LI-cadherin gene and protein in three groups, which showed:There were no obvious difference between control group and empty plasmid carrying group (P>0.05); The expression of LI-cadherin gene and protein were significantly lower than control group and empty plasmid carrying group, which had statistically significance (P<0.05)3. MTT assay was used to show that:The difference was no statistically significance among transfecting groups, control groups and blank groups in 12h,24h(P>0.05).The growth of transfecting groups was higher than control groups and blank groups in 36h,but the the difference was no statistically significance(P>0.05). The growth of transfecting groups'cells quickened in 48h and 72h, but the growth of control groups and blank groups change slowly. The growth rate of transfecting groups'cells was higher than control groups and blank groups, which had statistically significance (P<0.05)4. MTT assay was used to show that:The OD value and the number of three groups'cells respectively were 0.655±0.040,57.00±6.39;0.795±0.050,75.67±8.21; 0.821±0.026,79.33±4.32. The OD value and the number of transfecting groups' cells lower than control groups and blank groups, which had statistically significance (P＜0.01). The difference was no statistically significance between control groups and blank groups (P>0.05)5,Boyden chamber invasion assey showed that:The number of penetrating cells of three groups'cells respectively were:87.83±8.59;67.50±8.50; 68.33±7.50. The number of penetrating cells of transfecting groups'cells lower than control groups and blank groups, which had statistically significance (P<0.01). The difference was no statistically significance between control groups and blank groups (P>0.05)Conclusion:1. LI-cadherin-SiRNA was successful transfected to hepatocellular cells Hep3B.2. The transfection of LI-cadherin-SiRNA can suppress the expression of LI-cadherin gene and protein in Hep3B.3. The down of LI-cadherin can enhance the growth speed of the Hep3B; decrease the capacity of adhesion and migrant.