International Journal of Medical Sciences

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16 December 2018

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Int J Med Sci 2013; 10(10):1301-1306. doi:10.7150/ijms.6607

Research Paper

Lentivirus-based RNA Silencing of Nemo-like Kinase (NLK) Inhibits the CAL 27 Human Adenosquamos Carcinoma Cells Proliferation and Blocks G0/G1 Phase to S Phase

Bin Zhang1,2, Ke Yi Li2, Hai Ying Chen3, Shao Dong Pan3, Shuang Feng Chen3, Wei Feng Zhang2, Chun Peng Xia2, Li Cheng Jiang2, Xian Bin Liu2, Feng Jun Zhao3, Dao Ying Yuan2, Le Xin Wang2,4, Ya Ping Wu3,5✉, Shu Wei Liu1✉

1. Department of Anatomy Shandong University, School of Medicine, Jinan Shandong, 250012, P. R. China.
2. Department of Oral and Maxillofacial Surgery, Liaocheng People's Hospital, Liaocheng Shandong, 252000, P. R. China
3. Key Laboratory of Oral and Maxillofacial-Head and Neck Medicine, Central Laboratory, Liaocheng People's Hospital and Liaocheng Clinical School, Liaocheng Shandong, 252000, P. R. China.
4. School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW 2650, Australia
5. Department of Clinical Chemistry and Haematology, University Medical Centre Utrecht, G03.550, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands


Background: The Nemo-like kinase (NLK) is a serine/threonine-protein kinase that involved in a number of signaling pathways regulating cell fate. Variation of NLK has been shown to be associated with the risk of cancer. However, the function of NLK in oral adenosquamous carcinoma cells line CAL-27 is unknown.

Methods: In this study, we evaluated the function of NLK in CAL-27 cells by using lentivirus-mediated RNA silence. The targeted gene expression, cell proliferation and cell cycle are investigated by RT-PCR, western-blot, MTT method, colony forming assay and flow cytometry analysis respectively.

Results: After NLK silencing, the number of colonies was significantly reduced (54±5 colonies/well compared with 262±18 colonies/well in non-infected or 226±4 colonies/well in negative control group (sequence not related to NLK sequence with mismatched bases). Using crystal violet staining, we also found that the cell number per colony was dramatically reduced. The RNA silencing of NLK blocks the G0/G1 phase to S phase progression during the cell cycle.

Conclusions: These results suggest that NLK silencing by lentivirus-mediated RNA interference would be a potential therapeutic method to control oral squamous carcinoma growth.

Keywords: Nemo-like kinase (NLK), Lentivirus, RNAi, Oral Squamous Cell Carcinoma

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) License. See for full terms and conditions.
How to cite this article:
Zhang B, Li KY, Chen HY, Pan SD, Chen SF, Zhang WF, Xia CP, Jiang LC, Liu XB, Zhao FJ, Yuan DY, Wang LX, Wu YP, Liu SW. Lentivirus-based RNA Silencing of Nemo-like Kinase (NLK) Inhibits the CAL 27 Human Adenosquamos Carcinoma Cells Proliferation and Blocks G0/G1 Phase to S Phase. Int J Med Sci 2013; 10(10):1301-1306. doi:10.7150/ijms.6607. Available from