Expression And Significance Of MiR-103a-3p In Pancreatic Carcinoma

Objective：This study aims to detect the expression of miR-103a-3p in different cells.we selecttwo kinds of pancreatic cancer cell lines,one is PANC-1which has higher invasivenessthan BxPC-3(PANC-1>BxPC-3) and the normal pancreatic ductal epithelial cell line(H6C7).To verify the expression of miR-103a-3p in pancreatic cancer cells and pancreaticcarcinoma group whether the result is same or not. The next step is to select pancreaticcancer cell lines which have high expression of miR-103a-3p, constructe the lentivirusvector with the expression of miR-103a-3p what has been inhibited,then transfected theselected cell line, detected the expression of miR-103a-3p by qRT-PCR. This study mayprovide basis and reference of experimental and theoretical for the vivo tests andmechanism target genes.Methods:1. The expression of miR-103a-3p in PANC-1,BxPC-3and H6C7has been detected byqRT-PCR;2. Constructe the lentivirus vector with the expression of miR-103a-3p what has beeninhibited,After lentivirus Packaged,obtaind the supernatant of lentiviral particles;Transfect pancreatic cancer cell lines which have high expression of miR-103a-3p.3. Observe the transfection efficiency and detect its expression of miR-103a-3p byqRT-PCR.Results:1. QRT-PCR results showed that: the expression of miR-103a-3p in normal pancreaticcell lines (H6C7) and different invasion and metastasis of pancreatic cancer cell lines(PANC-1and BxPC-3) was determined. The2-ΔΔC(t)of miR-103a-3p is4.949±0.130inPANC-1cells; The2-ΔΔC(t)of miR-103a-3p is1.417±0.120in BxPC-3cells; The2-ΔΔC(t)of miR-103a-3p is1.010±0.151in H6C7cells。P<0.05was considered statisticallysignificant. The expression of miR-103a-3p in the three cells: PANC-1> BxPC-3> H6C7.2. Constructing miR-103a-3p-inhibitor lentiviral vector based on GV-159. After wepackaged the vector and transfected293T cells, we collected virus particlessupernatant titer and detected the viral titers about5×108TU/ml, infected pancreaticcancer cell lines. Then its transfection efficiency was observed. We obtained thestable expression of pancreatic cancer cell lines. The PANC-1cells which wereinfected with miR-103a-3p-inhibitor lentivirus were named as control-PANC-1,whichwere infected with negative lentivirus were named with NC-PANC-1. The results ofmiR-103a-3p in three cells (PANC-1、NC-PANC-1、control-PANC-1)：The2-ΔΔC(t)ofmiR-103a-3p is1.002±0.16in PANC-1cells; The2-ΔΔC(t)of miR-103a-3p is0.956±0.25in NC-PANC-1cells; The2-ΔΔC(t)of miR-103a-3p is0.317±0.32inControl-PANC-1cells。The PANC-1compared with NC-PANC-1, there is nostatistical significanc(P＞0.05), the control-PANC-1compared with PANC-1andNC-PANC-1respectively, the results were considered statstically significant.Conclusion：1. The expression of miR-103a-3p was significantly higher in pancreatic cancer celllines which has higher invasiveness.2. After transfected by lentivirus miR-103a-3p-inhibitor, its expression was significantlyreduced, and obtained the stably transfection PANC-1cell lines.