The Studies Of Interferences To The Expression Of Colon Cancer Cell Line’s Galectin-3Protein By SiRNA

Background:Small interfering RNA (siRNA) is a length of21-25nucleotides smallRNA molecules by Dicer (RNAase Ⅲ family having specificity fordouble-stranded RNA enzymes) processing, which is a principal member ofsiRISC and can stimulate a complementary target mRNA in silence. Previousstudies have shown that: galectin-3protein in colorectal cancer PhaseII solid tumor specimens showed a high expression of the trend,which isdetected by immunohistochemistry.Compared with adjacent tissues,thedifference is significant.Moreover,subsequent experiments found that lowmolecular weight citrus pectin (Low molecular weight citrus pectin, LCP)+oxaliplatin group can increase the apoptosis of colon cancer cellHCT-116and HT29.Compared with in the monotherapy group,the differencewas significant；In addition, Through a mouse model of colorectal livermetastases,it is found that: Galectin-3in colon cancer liver metastaseswas expressed at a higher levels compared with the negative control groupby ELISA, the difference is significant(P<0.05), and the LCP can inhibitthe occurrence of colorectal liver metastases.This experiment is not onlydesigned to confirm the roles of galectin-3protein in colorectal cancertreatment furtherly but also provide a active role for of the theoreticalbasis of targeted therapy of colorectal cancer.Methods: 1.Cell culture;2.The western bloting method to filter out the cell which express thehighest content of galectin-3protein;3.Close the expression of galectin-3with siRNA;4.Using the western bloting method to detect the effect of closing theexpression of galectin-3with siRNA;5.Using flow cytometry to detect the change of apoptosis;6.Statistical Methods:The IC50values are calculated by CompuSyn software.One-way ANOVA ort-test is used to compare the differences among the four groups by theSPSS13.0software.The inspection standard is а=0.05.Results:1.The galectin-3protein is detected by western bloting method in fivecolon cancer cell lines including HCT116、HT29、SW116、SW480and SW620,andWe use Brandscan5.0software to analysis the WB results.The expressionof galectin-3protein from high to low is SW11656.2%、HCT-11649.3%; HT-2946.2%、SW62043.6%and SW48046.2%；2.Two Gal-3specific shRNAs are used to interfer the expression ofSW116’s galectin-3protein by lentiviral vector pLVTHM.Through the grayanalysis,the interfering efficiencies is54.3%and61.9%,respectively；3.The apoptosis is detected by flow cytometry and the result is analysisedby SPSS13.0software.Compared with SW116cells, the difference issignificant (P <0.05).Conclusions:1. SW116has the highest expression of galectin-3protein in five coloncancer cell lines including HCT116、HT29、SW116、SW480and SW620；2.The interference effect targeting on two Gal-3specific shRNAs bylentiviral vector pLVTHM is54.3%and61.9%, respectively；3.Oxaliplatin can increase the interfered SW116cell’s apoptosis. Compared with SW116cells, the difference is significant (P <0.05).