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The effect of ASPM gene in the acute damage induced by Cadmium chloride for 16HBE cells

Shanshan Z  Liang R  Piaofan Y  Xiaoxuan Y  Ping Y  Yixiong L  
【摘要】:Objective This studyattempts to investigate the relationships between ASPM role and lung tumor Cadmium caused, which might be a new biomarker for diagnosis, treatmentand prognosis of lung cancer.Methods The expression of ASPM at m RNA and protein level in 16HBE cells under CdCl2 treatment was detedcted by qRT-PCR and immunohistochemical analysis.ASPM low-expression cell lines were constructed by hairpin RNA(shR-NA) for obseving cell changes which were treated with CdCl2. 16HBE cells were treated respectively with control,10μM/L, 20μM/L, 30μM/L and 40μM/L CdCl2 for 24h, and then 16HBE cellswere treated with30μM/L CdCl2 for 12h, 24h, 48h and 72h. The function of ASPM gene was detected by MTT,comet assay, cell flow technology and Cytochrome c apoptosis assay. Subsequently, these datas were statistically analyzed.Results With the dose and time increase of CdCl2, the expression of ASPM upregulated in 16HBE cells. Compared with 16HBE cells, there was no statistical differences of growth and morphlogy in cells proliferation of ASPM dificiency. Our data, from the Comet assay in detail, indicated that cadmium chloride caused DNA damage in 16HBE cells in a concentration-time dependent manner, yet the seriousness of DNA damage was more in 16HBEshASPM cells. In H2O2-induced oxidative damage, the DNA damage, for down-regulating the expression of ASPM, was also more than control group. In MTT, ASPM gene can be related to decreasing cell viability in the process of cell damage induced by Cadmium chloride. With theincrease of concentration and time, the release of cytochrome C increased. In 16HBE-shASPM cell, the release of cytochrome C was higher than 16HBE cells at the identical procedure. It indicated that ASPM gene under-expression could increase apoptosis and release cytochrome C.Conclusions Abnormal spindle microtubule assembly (ASPM) gene was regarded as a biomarkers for diverse tumors. However, the research has not been particularly obvious in regarded to carcinogenic mechanism of ASPM about lung neoplasms cadmium-caused. Our data suggests that ASPM will play a role in 16HBE cells damaged by Cadmium chloride. ASPM will play a positive effect on cell damaged by cadmium. Above all, this study provides a clue that there is a link between ASPM gene and lung neoplasm cadmium-caused.

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