Found programs:
Authors:Wang Huimin; Cui Can; Wang Yinxin; Li Yanfeng; Yuan Dongjie; Lu Zhenmin
Keywords:laryngeal carcinoma;transient receptor potential M7;oxidative stress;proliferation;apoptosis;invasion
DOI:10.19405/j.cnki.issn1000-1492.2022.05.007
〔Abstract〕 Objective To explore the effects and mechanisms of interfering with transient receptor potential melastatin subfamily member 7(TRPM7) on biological behaviors(proliferation, apoptosis, invasion) of laryngeal carcinoma cells. Methods Human laryngeal carcinoma cells TU212 were culturedin vitro. TRPM7-shRNA plasmid vectors(TRPM7-shRNA1 group, TRPM7-shRNA2 group, TRPM7-shRNA3 group) and negative control shRNA-NC(shRNA-NC group) were constructed. TU212 cells were transfected by liposome transfection method. The cells transfected with empty vector were enrolled as Control group. The level of lactic dehydrogenase(LDH) in TU212 supernatant was detected by ELISA. The level of malondialdehyde(MDA) and activity of superoxide dismutase(SOD) in supernatant were detected by colorimetry. The effects of knocking-down TRPM7 on proliferation and invasion of TU212 cells were detected by CCK-8, clone formation assay and Transwell assay. The expressions of invasion and apoptosis-related proteins were detected by Western blot. Results After transfection, expression levels of TRPM7 mRNA and protein were down-regulated in TRPM7-shRNA1 group, TRPM7-shRNA2 group and TRPM7-shRNA3 group compared with Control group(P<0.05), and the decrease was the most significant in TRPM7-shRNA1 group(P<0.05). In functional experiments, SOD level in TRPM7-shRNA1 group decreased compared with Control group(P<0.05), while MDA and LDH levels increased(P<0.05). The cells proliferation rate and clone formation rate were decreased(P<0.05), the number of invasion cells was reduced(P<0.05), the expressions of N-cadherin and Vimentin proteins were down-regulated(P<0.05), mitochondrial membrane potentials were reduced(P<0.05), Bax/Bcl-2 and cleaved caspase-3/caspase-3 increased(P<0.05). Conclusion Knocking-down TRPM7 can increase oxidative stress level in laryngeal carcinoma cells TU212, inhibit their proliferation and invasion, and induce their apoptosis by mitochondrial pathways.