The Indentification and Suppression Effection of Proliferation on Human Laryngeal Cancer Stem Cell
Wang Binrong, Ying Chang, Fang Huiling, Wang Yigang*
The incidence of laryngeal cancer is increasing year by year. Traditional chemotherapy drugs such as cisplatin, doxorubicin hydrochloride, are essential in killing laryngeal cancer cells, reducing laryngeal cancer distant metastasis and improving patient survival, but they are not effective in killing cancer stem cells (CSCs). Targeting or selectively killing CSCs is expected to improve the sensitivity of chemoradiation, reduce cancer metastasis and recurrence. In this paper, laryngeal cancer stem cells (Hep2 sphere) were obtained by suspension culture of Hep2 in vitro. The expressions of Hep2 sphere related factors, cell cycle and resistance were detected by qRT-PCR and Western blot, indicating that they have tumor stem cells characteristic to some extent. Cloning formation experiment and cell counting show that many Hep2 sphere are in the resting stste G0/G1 phase and their proliferation ability is weak. qRT-PCR results indicated Hep2 sphere upregulated CD44, ALDH1, p-AKT, bcl2 as well as p21 gene and downregulated bax, AKT gene campared to Hep2 cells. Western blot results confirmed that bax and AKT protein expression was downregulted in Hep2 sphere, bcl2 and p21 protein expression were significantly upregulated. MTT assay showed that the cell viability of Hep2 sphere was significantly higher than that of Hep2 under different drug treatments. Under the treatment of adenovirus ZD55-Trail, the cell survival rate of Hep2 was significantly higher than that of Hep2 sphere. This study have shown that suspension cultured laryngeal cancer stem cells have stem cell characteristics; adenovirus ZD55-Trail has stronger killing effect on laryngeal cancer stem cells, and weaker on differentiated Hep2, but DDP, Dox and 5-Fu are on the contrary.
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