FAKsiRNA Inhibits Hypoxia-induced Human Pulmonary Arterial Smooth Muscle Cells Proliferation By Down Regulation Of MTERF1 And CyclinD1

Objecive:To explore Whether FAKsi RNA inhibits the proliferation of human pulmonary artery smooth cell under hypoxia,via m TERF1/ Cyclin D1 signal pathway.Mehtods:Cultured primary HPASMCs were transtected by Lipofectamine2000 with the chemical synthetic human FAK,MTERF1 of si RNA. This cells are grouped into nomoxic(37℃,5%CO2,21%O2)Noncontrol si RNA transfecting group,nomoxic FAKsi RNA transfecting group,nomoxic m TERF1 si RNA transfecting group; hypoxic( 37℃,5%CO2,5%O2,90%N2) Noncontrol si RNA transfecting group,hypoxic FAKsi RNA transfecting group,hypoxic m TERF1 si RNA transfecting group. Use Realtime-PCR detectes the expression of FAKm RNA,MTERF1 m RNA and Cyclin D1 m RNA level,;Western Blot and immunofluorescence detect the protein change of FAK, Cyclin D1;CCK8 methods was observed for the effection of cell proliferation;Results: Compared with normoxic NCsi RNA group, Both FAKm RNA and protein expression were significantly reduced in normoxic FAKsi RNA group(P <0.05), whereas there were no significant difference of MTERF1 and Cyclin D1,(P > 0.05); compared with hypoxia NCsi RNA group, FAKm RNA and protein expression was significantly reduced in hypoxia.FAKsi RNA group(P <0.05), subseqently, the protein and m RNA of m TERF1 and Cyclin D1 were decreased(P <0.05); After transfected m TERF1 si RNA,Comparing with normoxic NCsi RNA group.the expression of m TERF1 m RNA and protein were significantly reduced in normoxic m TERF1 si RNA group(P <0.05), whereas both FAK and Cyclin D1 of m RNA and protein had no significant change(P>0.05). Compared with the hypoxia group NCsi RNA, m TERF1 m RNA and protein expression were significantly reduced in hypoxia m TERF1 si RNA group(P <0.05), however,Cyclin D1 m RNA and protein expression were significantly reduced in hypoxia m TERF1 si RNA group(P <0.05), while FAKm RNA and protein expression were no significant change(P>0.05) in hypoxia m TERF1 si RNA group. Detection of CCK8 displays that regardless of under normoxic or hypoxic conditions, compared with Noncontrolsi RNA,the proliferative activity of FAKsi RNA group and m TERF1 si RNA group were significantly lowered.(P<0.05) Immunofluorescence assay shows that the fluorescence expression of FAK was significantly decreased in FAKsi RNA group comparing to normoxia NCsi RNA group(P <0.05), In like manner Cyclin D1 was no significant change(P> 0.05).however in hypoxia, compared with NCsi RNA group, the fluorescence expression of FAK was significantly decreased in FAKsi RNA group(P <0.05), so did the Cyclin D1(P <0.05).Conclusion : Under Hypoxia, FAK gene knockouted, can not only reduce the expression of m TERF1 and Cyclin D1 but also inhibit the proliferation of HPASMCs. knocked down m TERF1 gene,the expression of Cyclin D1 reduced, However,the expression of FAK protein and m RNA have no significant change. which suggests m TERF1/ Cyclin D1 be situated on the downstream signaling pathways of FAK.The fact illuminates FAK-m TERF1/ Cyclin D1 participate in the proliferation of HPASMCs in hypoxia.