Construction of Lentiviral Vector of Naa10 Small-interfering RNA and Its Effect on Growth of Oral Squamous Cell Carcinoma.

doi:10.13701/j.cnki.kqyxyj.2017.07.005

Abstract

Abstract: Objective: To construct the effective lentiviral vector of Naa10 small-interfering RNA, and transfect it into oral squamous cell carcinoma SCC-15 cells so as to investigate its effect on the growth of SCC-15 cells. Methods: Three pairs of siRNA sequences against different parts of Naa10 mRNA were separately transfected into SCC-15 cells. The transfection efficiency was verified, and then the most effective shNaa10 sequence was cloned into pLV-shRNA. pLV-shNaa10 was verified by sequencing and co-transfected into 293T cells to obtain packaged lentivirus particles LV-shNaa10. After viral titer determination, SCC-15 cells were infected with LV-shNaa10 and the expression of Naa10p was detected by Western Blot. Growth curves were used to study the effect of Naa10 knockdown on the growth of SCC-15 cells. Results: Naa10 siRNAs could suppress Naa10 expression and the second siRNA was the most effective (P<0.005). LV-shNaa10 and LV-NC harvested from 293T cells had a titer of 1.2×10⁹ TU/mL and 1.0×10⁹ TU/mL, respectively. After infection with LV-shNaa10, the expression of Naa10 protein in SCC-15 cells was down-regulated (P<0.005), which could in turn markedly facilitate the growth of SCC-15 cells. Conclusion: The lentivirus-mediated Naa10 shRNA was successfully constructed, and knocking down of Naa10 could promote the growth of SCC-15 cells.

Key words: Naa10 , Lentivirus, RNA interference, Oral squamous cell carcinoma

CLC Number:

R739.8

YANG Yang, ZHENG Jun, XU Jiang, GU Yong-qing, HUANG Jin, WANG Dan-ni, ZHU Mao-xiang, ZENG Yan.. Construction of Lentiviral Vector of Naa10 Small-interfering RNA and Its Effect on Growth of Oral Squamous Cell Carcinoma.[J]. Journal of Oral Science Research, 2017, 33(7): 707-711.

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