Abstract
Objective To observe the effect of ORAI1 on cell migration and invasion in prostate cancer PC-3 cells, and explore whether ORAI1 contribute to reverse the epithelial-mesenchymal transition (EMT) transformation. Methods PC-3 cells were transfected with lentiviral vector LV-ORAI1-RNAi (KD group) and related negative vector (NC group), the transforming growth factor-β1 (TGF-β1) was added separately. After transfection, the expression of ORAI1 in PC-3 cells was detected by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blot. The cell mobility, migration and invasion ability of PC-3 cells were measured by Celigo scratch assay, transwell migration and invasion assays, respectively. The expression of EMT markers [E-cadherin, N-cadherin, Vimentin], and smad3, p-smad3 were detected by Western blot analysis. Results After transfected with LV-ORAI1-RNAi, ORAI1 expression in PC-3 cells was decreased at both mRNA and protein levels(P<0.05). Celigo scratch assay showed that the mobility of KD group was (10.24%±1.17%) decreased (P<0.05) compared with that of NC group (14.38%±0.77%), the mobility of KD+ TGF-β1 group (12.44%±1.29%) was decreased (P<0.05) compared with that of NC+ TGF-β1 group (26.21%±2.23%). Transwell migration assay showed that the number of migrating cells in KD group (229+ 8.4) was lower than that in NC group (298+ 14.2) (P<0.05); compared with NC+ TGF-β1 group (338+ 12.9), the number of migrating cells in KD+ TGF-β1 group (216+ 12.2) was lower (P<0.05). Transwell invasion assay showed that the number of invasive cells in KD group (46+ 3.4) was lower than that in NC group (79+ 4.6) (P<0.05); compared with NC+ TGF-β1 group (123+ 5.1), the number of invasive cells in KD+ TGF-β1 group (66+ 5.5) was lower (P<0.05). Western blot results showed that the expression of E-cadherin decreased, while the expression of N-cadherin and Vimentin increased in NC+ TGF-β1 group, compared with NC group. Compared with NC+ TGF-β1 group, the expression of E-cadherin increased, while the expression of N-cadherin, Vimentin and p-smad3 decreased in KD+ TGF-β1 group. Conclusions The decreased expression of ORAI1 in PC-3 cells may be related to the reversal of EMT through TGF-β/smad3 signaling pathway. Key words: Prostate cancer; ORAI1; Tumor metastasis; Epithelial-mesenchymal transition
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