Effect Of STIM1in The Proliferation Of Non-small Lung Cancer

Objective:Ca2+is known as one of the second signaling factor in cells, and the Ca2+concentration may affect cellular functions, such as cell proliferation, mobile and gene expression. STIM1is the Ca2+sensor, which is located in the endoplasmic reticulum. STIM1is capable of sensing the concentration of Ca2+in ER and trigger the activation of SOC. Preliminary studies suggest that STIM1is closely related to a variety of cell proliferation. The aim of current study is to confirm that STIM1would play an important role in proliferation of NSCLC cells, which would provide scientific basis for a further understanding of the molecular mechanisms in the progression of NSCLC.Methods:●Expression of STIM1in H522,H2405,H2342,A549,SPC-A-1,SW900, H1869,SK-MES-1, H1299,H661, H1581was measured by Western blotting.●Among all the NSCLC cell lines, A549cell line was infected by STIM1-shRNA lentiviral vector as experimental model. RT-PCR and Western blotting were performed to detect the mRNA and protein expression of STIM1after infection.●MTS cell proliferation assay, flow cytometry cell cycle analysis were used to analyze the alterations of NSCLC biological behaviors.●By immunochemical SP techenique, we detected the expression of STIM1in the pathological paraffin tissues of NSCLC and benign lesions, then analyzed the association between them by SPSS software.●BALB/c nude mice were used to detect the effect of STIM1during the tumor formation in vivo.●Use RT-PCR to analyze the proliferation-related genes after knockdown of STIM1.Results:●Western blotting results showed that STIM1is stable expressed in all NSCLC cell lines (H522,H2405,H2342,A549,SPC-A-1,SW900,H1869,SK-MES-1, H1299,H661,H1581).●RT-PCR and Western blotting results showed that endogenous STIM1was silenced by STIM1-shRNA lentiviral vector.●Knockdown of STIM1inhibited NSCLC cells proliferation and caused cell cycle arrest at G1phase.●The expression rate of STIM1is significantly higher in the pathological paraffin tissues of NSCLC (79.83%) than in the in benign lesions (18.72%)(P<0.05). And STIM1expression was higher in squamous cell carcinoma than in adenocarcinoma (P<0.05).●STIM1-shRNA induced significant retardation of tumor growth in nude mice.●RT-PCR results revealed that CDK2,CDK9,CDK9and cyclinD3were decreased after knockdown of STIM1. Conclusions:Aberrant expression of STIM1might participate in the malignant progression of NSCLC.