Effects Of Targeted Silence Adrenomedullin Gene On Invasion And Migration Of The Pancreatic Cancer Cell Line MiaPaCa-2

Objectives:To investigate the effect of RNA interference(RNAi) plasmid targeted silence ADM gene on the migration and invasive ability of pancreatic cancer cell line MiaPaCa-2and its action mechanism. And this study may offer a new approach for pancreatic cancer clinical therapy.Methods:â‘ Pancreatic cancer cell line MiaPaCa-2were cultivated.The expressions of ADMmRNA were measured by PCR qualitatively; the expressions of ADM proteins were measured by western blotting qualitatively.â‘¡To design and synthesize the experimental group plasmid PgenesilADM and the negative control group PgenesilHK. PgenesilADM contains two target shRNA,and PgenesilHK has a non-specific sequence.Then, to identify and purify the plasmids.â‘¢The pancreatic cancer cells were intervened by PgenesilADM,PgenesilHK and the empty plasmid PgenesilKB:The cells were randomly divided into control group,PgenesilKB intervention group,PgenesilHK intervention group and PgenesilADM intervention group. After transfection for48hours, transwell invasion and migration test were given to examine the changes of the migration and invasive ability of The pancreatic cancer cells.â‘£After transfection for48hours, the expressions of ADMmRNA were measured by real-time fluorescence quantitative PCR; the expressions of ADM proteins were measured by Western Blot method.Results:â‘ By PCR and Western Blot, ADM mRNA and ADM proteins were expressed within the pancreatic cancer cell line MiaPaCa-2.â‘¡The migration and invasive ability of pancreatic cancer cells in PgenesilADM intervention group was significantly lower than that in control group (P<0.05),PgenesilKB intervention group (P<0.05),PgenesilHK intervention group (P<0.05).But there was no difference between the control group,PgenesilKB intervention group and PgenesilHK intervention group (P>0.05).â‘¢By real-time fluorescence quantitative PCR,the levels of ADMmRNA expression of pancreatic cancer cells in PgenesilADM intervention group was significantly lower than that in control group (P <0.05),PgenesilKB intervention group (P<0.05)and PgenesilHK intervention group (P<0.05). But there was no difference between the control group,PgenesilKB intervention group and PgenesilHK intervention group (P>0.05).â‘£By Western Blot,the levels of ADM proteins expression of the pancreatic cancer cell line MiaPaCa-2in PgenesilADM intervention group was significantly lower than that in control group (P<0.05),PgenesilKB intervention group (P<0.05)and PgenesilHK intervention group (P<0.05). But there was no difference between the control group,PgenesilKB intervention group and PgenesilHK intervention group (P>0.05).Conclusions:1. ADMmRNA and proteins were expressed in the pancreatic cancer cell line MiaPaCa-2.2. The recombinant plasmid PgenesilADM transfects pancreatic cancer cell line MiaPaCa-2can effectively reduce the migration and invasive ability of pancreatic cancer cells, but its specific mechanism needs further research.3. RNAi technology targeted silence ADM gene can reduce the expression of ADM mRNA and proteins in pancreatic cancer cell line MiaPaCa-2. This suggests that the decrease expression of ADM may contribute to reduce migration and invasive ability of pancreatic cancer cell line MiaPaCa-2, but the specific molecular mechanism remains to be further research.