The Expression Of RhoC And RhoGDIα In Lung Carcinoma Cells And Their Correlation With The Ablility Of Invasion

INTRODUCTIONRho protein family is one member of Small GTP-binding proteins of Rassuperthmily. RhoC is one member of Rho subtribe, which take a pivotal role in theprocess of tumor invasion and metastasis. However, there is no studies about RhoCexpression in lung carcinoma and its correlation with metastasis of lung cancer.RhoGDIs have been regarded as regulators to inactivate Rho proteins, however,evidence is accumulating that the classical function of rhoGDIs as universal chaperonesfor GDP-bound proteins may need to be revisited. Several studies have investigated thelevel of expression of rhoGDIs in various cancer cells compared with normal cells,revealing opposite patterns depending on the tumor cell considered. Likewise, rhoGDIβmRNA levels were increased in ovarian adenocarcinoma, and this upregulationcorrelated with the malignancy of tumors. In addition, the induction of motility ofcancer cells by the autocrine motility factor cytokine appeared to induce anupregulation of rhoGDIβmRNA and protein in vitro. However, less studies reportedthe correlation of Rho-GDIαexpression and metastasis of cancer. In the present study,We investigate the expression of RhoC and Rho-GDIαmRNA and protein in four lungcarcinoma cell (BE1, LH7, A2, A549) and Homo sapien Bronchus Epithelial cell(HBE)and their correlation with metastasis of lung cancer. METERAL AND METHODS1. Material and ReagentsHuman lung cancer cells BE1, LHT, A549, A2 were obtained from Beijing MedicalUniversity. Homo sapien bronchusepithelial cell was purchased from ZhongnanUniversity Xiangya laboratory. RPMI 1640 were purchased from Gibco. The firstantibodies were rabbit polyclonal antibody to RhoC and goat polyclonal antibody toRho-GDIαwas purchased from Santa Cruz. Trizol was purchased from Tiangen.2. MethodsThe expressions of RhoC, Rho-GDIαprotein were detected with western blot.The expressions of RhoC, Rho-GDIαRNA were detected with reverse-transcription polymerase chain reaction (RT-PCR).The expressions and locations of RhoC, Rho-GDIαwere detected with immuno-fluorescence.The invasion ability of BE1 and LH7 in vitro was detected with BioCoat MatrigelInvasion Chambers with 8.0 micron pore transwells coated with extracellular matrixproteins.3. Statistical analysisThe SPSS 11.5 software was employed to ayalyze the data. P＜0.05 wasconsidered as statistical significance.RESULTSThe expression level of RhoC and Rho-GDIαprotein in various lung carcinomacell was significantly higher than that in HBE. The expression of RhoC and Rho-GDIαprotein in BE1(the cell with high metastasis capability) was significantly higher thanthat in LH7(the cell with low metastasis capability). The expression of RhoC andRho-GDIαmRNA in BE1(the cell with high metastasis capability) was significantlyhigher than that in LH7(the cell with low metastasis capability). The invasion ability ofBE1 in vitro was significantly higher than LH7. The expression of RhoC and Rho-GDIαin BE1 and LH7 was all located in endochylema.CONCLUSIONThe expression level of RhoC and Rho-GDIαin lung carcinoma cell was higherthan in HBE.The expression of RhoC and Rho-GDIαcorrelates with metastasis of lung cancer.