ABSTRACT
Objective To investigate the underlying mechnisms of Kangai-1(KAI1)gene,a tumor metastasis suppressor gene,on metastasis and prolification of pancreatic cancer cells.Methods The plasmin containing Ad-KAI1 was established and transfected into pancreatic cancer cell line PCNA Ⅰ.The PCNA Ⅰ cells were then divided into different groups according to the times induced by vascular endothelial growth factor(VEGF)and epidermal growth factor(EGF).The morphology and migrational ability of PANC Ⅰ cells were compared before and after transfection by microscopy and transwell method,respectively.The expressions of intercellular adhesion molecule-1(ICAM-1)and matrix metalloproteinases-9(MMP-9)in PANC Ⅰ cells were examined by immunocytochemistry before and after transfection.Results The migrational ability of PCNA Ⅰ cells transfected with Ad-KAI1 was siginificantly decreased compared with untransfected PCNA Ⅰ cells(P＜0.05).Immunocytochemistry study revealed that the expressions of ICAM-1 and MMP-9 were both positive in untransfected PCNA Ⅰcells,but were both negative in transfected PCNA Ⅰ cells.Conclusion The inhibitory mechanism of KAI1 gene on metastasis of pancreatic cancer is associated with down-regulation of ICAM-1 and MMP-9expressions.
ABSTRACT
Objective To evaluate the expression of KAII (CD82) gene inhibited by small interfering RNA (siRNA) in human pancreatic cancer cell line T3. Methods Four sequences of siRNA including A, B,C, D were designed, which were based on the KAI1 gene sequence using online RNA interfering designing software and lentivirus vector was built. Then they were used to transfect T3 cells by liposome 2000 and virus titer was determined. Empty vector containing siRNAd1 lentivrus particle ( MOI =5) was also used to infect T3 cells. The expression of CD82 mRNA was detected by real-time PCR. Results The expression of CD82 mRNA in normal control group, empty vector group, A group, B group, C group, D group were 1. 398 ±0.242,1. 311±0.048, 0. 664 + 0. 093, 0. 345 ± 0. 032, 0. 641 ± 0. 049 and 0. 147 ± 0. 049, respectively, the difference between the expression of CD82 mRNA in empty vector group and that of A, B, C, D groups was significant (P＜0.01 ). Conclusions RNAi was able to inhibit the expression of KAI1 gene CD82 in human pancreatic cancer cell line T3.