Influence of nutrient-deprivation autophagy factor-1 gene on the proliferation and metastasis behavior of human tongue cancer cells

doi:10.3877/cma.j.issn.1674-1366.2014.06.003

Abstract

Abstract:

Objective

To investigate the inhibitory effect of small interfering RNA （siRNA） on the expression of nutrient-deprivation autophagy factor-1 （NAF1） in human tongue cancer Tca8113 cell line via transfected with the special siRNA. The biological behaviors， including cell proliferation and metastasis of the transduced cells were investigated as well.

Methods

The cultured Tca8113 cells were divided into three groups including si-NAF1 group， si-NC group and the vector group. MTT assay and plate cloning formation assay were used to identify the effect of NAF1-siRNA on the cell proliferation.Transwell assay.was used to investigate the invasion capability of the Tca8113 cells. The RNA and protein levels of functional biomarkers such as cyclin D1 and MMP-2 were detected through RT-PCR and Western blot assay.

Results

NAF1 expression in Tca8113 cells transfected with NAF1-siRNA was significantly down regulated at both the RNA and protein levels compared with those in normal Tca8113 cells （χ²=25.65，t=-17.1，P ＜0.05）， accompany with the decreases of cyclin D1 and MMP-2 protein（t_cyclin_D1=-14.7，t_MMP-2=-9.6，P ＜0.05）. The proliferation and cloning formation of Tca8113 cells were obviously inhibited in si-NAF1 group compared with the vector group （t=-36.77，t=-12.33，P＜0.05）. The cell invasion of Tca8113 cells was obviously inhibited in si-NAF1 group compared with the vector group.

Conclusion

Down regulation of NAF1 inhibited tongue cancer cells proliferation and metastasis， which might mediated via cyclin D1 and MMP-2 protein.

Key words: Tongue neoplasms, Neoplasms， squamous cell, RNA interference, Nutrientdeprivation autophagy factor-1deprivation autophagy factor-1

Guohua Jiao, Zhihui Kan, Hongli Gao, Zhiguang Zhang. Influence of nutrient-deprivation autophagy factor-1 gene on the proliferation and metastasis behavior of human tongue cancer cells[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2014, 8(06): 454-458.

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URL: https://zhkqyxyjzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-1366.2014.06.003

https://zhkqyxyjzz.cma-cmc.com.cn/EN/Y2014/V8/I06/454

Figures/Tables 8

References 26

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基因	引物序列
NAF1	F：5'-ATGCTTTTTCATCATGTTCAACT-3'
	R：5'-GCATCAGGGCTTTCAATGCAC-3'
β-actin	F：5'-TCAATGATTTGGCTACAGCA-3'
	R：5'-AGATCATGTACATGGCAACTG-3'

基因	引物序列
NAF1	F：5'-ATGCTTTTTCATCATGTTCAACT-3'
	R：5'-GCATCAGGGCTTTCAATGCAC-3'
β-actin	F：5'-TCAATGATTTGGCTACAGCA-3'
	R：5'-AGATCATGTACATGGCAACTG-3'

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