The Relationship Of ORAI1 On The Biology Characteristics Of Colon Cancer

Objective:To explore the effect of ORAI1 on the proliferation and metastasis of colon cancer cell line SW480 and its mechanism, RNA interference was used to inhibit the gene expression of ORAI1.Methods:The SW480 cells was infected with recombinant lentivirus, there are control group, empty virus group, interference group in the experiment. The expression of ORAI1 mRNA and protein was confirmed by fluorescence quantitative PCR(FQ-PCR) and Western blotting. MTT、 colony-forming and doubling time was used to examined the proliferation of SW480, Transwell chamber, adhesion,angiogenesis and vasculogenic mimicry experiments were used to detect the ability of invasion, migration, angiogenesis, homogeneous and heterogeneous cells intercellular adhesion among each group, flow cytometry was used to detect the periodic variation of SW480, confocal microscopy was used to detect the difference of SOCE (store-operated Ca2+ entry, SOCE) in three groups of cells. Meanwhile, Western blotting was used to detect the expressions of ERK1/2、p-ERK1/2、CyclinD1、MMP-2 VEGF and E-cadherin protein.Results:After infected SW480 with ORAI1 interference lentivirus for 72h,significant fluorescence expression can be seen. Compared with the empty vector group and the control group,the expression of ORAI1 was lower (P<0.01),the cells grew slower(P<0.05),the colony-forming ability was decreased (P<0.01), the doubling time was delay(P<0.01), the invasion and migration ability were diminished (P<0.01), the homogeneous cells intercellular adhesion ability were incresed (P<0.05), the heterogeneous cells intercellular adhesion ability were decreased (P<0.05), the angiogenesis and vasculogenic mimicry were enhanced(P<0.01), the proportion of cells in G1 phase was higher(P<0.05), the Internal flow peak of SOCE was lower(P<0.05), and the expressions of p-ERK1/2, CyclinD1, MMP-2 and VEGF proteins were deseased(P<0.01), and the expressions of, the expressions of E-cadherin protein was incresed (P<0.01).Conclusion:ORAI1 may promote the proliferation and metastasis of SW480 colon cancer cell line, the mechanism may be associated with the increased of SOCE.