Effect of silencing calcium channel-related gene CACNA1A on proliferation, invasion and migration ability of human osteosarcoma cells
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Abstract
Objective: To investigate the effect of silencing calcium (Ca2+) channel-related gene CACNA1A on proliferation, invasion and migration of osteosarcoma (OS) cells. Methods: Reverse transcription-quantitative PCR (RT-qPCR) was used to detect the expression of CACNA1A in tumor tissues and adjacent tissues of clinical patients with OS. The expression of CACNA1A in osteoblasts (hFOB1.19 cells) and OS cell lines (MNNG and SJSA-1 cells) was compared. The small interfering RNA (siCACNA1A) knocking out CACNA1A and its control (siControl) were transferred into MNNG and SJSA-1 cells respectively to observe the Ca2+ distribution in the OS cells after silencing CACNA1A expression. Cell counting kit-8 (CCK-8) assay, scratch assay and Transwell assay were used to detect the cell proliferation, invasion and migration. A subcutaneous transplanted tumor model was established in nude mice, the volume and weight of tumorigenesis in the siCACNA1A group and the siControl group were compared, and the expression level of CACNA1A protein in tumorigenic tissues was detected by immunohistochemical staining (IHC). Results: Compared with adjacent tissues, the expression level of CACNA1A in tumor tissues of OS patients was increased (P<0.05). Compared with hFOB1.19 cells, the expression level of CACNA1A in MNNG and SJSA-1 cells was increased (P<0.05). Compared with the siControl group, the Ca2+ distribution in the OS cells in the siCACNA1A group was reduced, and the cell proliferation and migration ability was significantly inhibited. The results of animal experiments showed that the tumor volume and weight in the siCACNA1A group were significantly reduced compared with the siControl group (P<0.05). IHC results showed that the relative immunostaining intensity of CACNA1A in subcutaneous tumors in the siCACNA1A group was lower than that in the siControl group (P<0.001). Conclusion: The silencing Ca2+ channel-related gene CACNA1A can inhibit the proliferation and metastasis of OS cells and the growth of tumors in nude mice.
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