Effects of specific interfering TACC3 gene expression on proliferation and apoptosis of CD133+CD44+ oral squamous cell carcinoma cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Studies have indicated that the abnormal expression of TACC3 is closely related to the occurrence and development of many kinds of tumors, and the expression of TACC3 is up-regulated in these tumors. Therefore, in vitro specific inhibition of TACC3 expression may become an important target for the treatment or intervention of tumor growth.OBJECTIVE: To investigate the mechanism by which TACC3 gene expression regulates cell proliferation and apoptosis in oral squamous cell carcinoma.METHODS: CD133+CD44+ oral squamous cell carcinoma cells were sorted from human oral squamous cell carcinoma cell line Cal-27 by immunomagnetic beads. In experimental group, the shRNA sequence of TACC3 was designed and synthesized, which was then trasnfected into CD133+CD44+ oral cancer stem cells by LipofectamineTM 2000. Empty vector-trasnfected (negative control) and untransfected cells were used as callsed. Forty-eight hours after the transfection, effects of TACC3 gene silencing on proliferation and apoptosis in vitro in CD133+CD44+ oral squamous cell carcinoma were detected by MTT, clone formation test, and TUNEL assay. Western blot assay was used to detect the effect of TACC3 gene silencing on Ki67, Bax and Bcl-2 protein expression in CD133+CD44+ oral squamous cell carcinoma.RESULTS AND CONCLUSION: (1) Cell proliferation. The proliferation rate and expression level of Ki67 were significantly lower in the experimental group than the negative control and untransfected groups (P < 0.05). (2) Clone formation. The clone formation ability in the experimental group was significantly lower than that in the negative control and untransfected groups (P < 0.05). (3) Cell apoptosis. TACC3 gene silencing caused an obvious decrease in Bcl-2 protein expression and a significant increase in Bax protein expression. These findings further confirmed that specific interference of TACC3 gene expression could inhibit the proliferation of CD133+CD44+ cells and promote the apoptosis.