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Lentivirus‑delivered nemo‑like kinase small interfering RNA inhibits laryngeal cancer cell proliferation in vitro.

Tai J, Rao Y, Fang J, Huang Z, Yu Z, Chen X, Zhou W, Xiao X, Long T, Han Y, Liu Q, Li A, Ni X - Mol Med Rep (2015)

Bottom Line: The effects of NLK downregulation on Hep‑2 cell proliferation and cell cycle progression were analyzed using an MTT assay and flow cytometry, respectively.Downregulation of NLK also inhibited tumorigenesis and regulated the expression of cell cycle protein expression levels.Therefore, it was hypothesized that NLK is necessary for cell survival and tumorigenesis in laryngeal cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, Beijing 100045, P.R. China.

ABSTRACT
Laryngeal squamous cell carcinoma is the most common form of head and neck squamous cell carcinoma. Multiple approaches have been applied to treat this type of cancer; however, no significant improvement in survival rate has been achieved. In the present study, the role of nemo‑like kinase (NLK) in human laryngeal carcinoma Hep‑2 cells was investigated. NLK has been identified as an important regulator of cell growth, patterning and cell death in a variety of organisms. Lentivirus‑mediated‑shRNA was employed to silence endogenous NLK expression. Downregulation of the expression of NLK following lentivirus infection was confirmed using reverse transcription quantitative polymerase chain reaction and western blot analysis. The effects of NLK downregulation on Hep‑2 cell proliferation and cell cycle progression were analyzed using an MTT assay and flow cytometry, respectively. Downregulation of NLK also inhibited tumorigenesis and regulated the expression of cell cycle protein expression levels. Therefore, it was hypothesized that NLK is necessary for cell survival and tumorigenesis in laryngeal cancer cells. Furthermore, the absence of NLK may lead to cancer cell death. Collectively, the results of the present study demonstrated that the lentivirus‑mediated targeted disruption of NLK may be a promising therapeutic method for the treatment of laryngeal cancer.

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Effects of NLK on the cell cycle and cell apoptosis determined by flow cytometric analysis. (A) Histograms showing the cell cycle distribution of uninfected Hep-2 cells (Con) and cells infected with the Lv-shNLK and Lv-shCon. (B) Suppression of NLK expression by RNA interference resulted in an increase in the percentage of cells in the S phase. (C) A significant increase was identified in the percentage of cells in the sub-G1 phase following transfection with NLK siRNA. The percentage of cells in each phase of the cycle is presented as the mean ± standard error of three independent experiments. NLK, nemo-like kinase.
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f3-mmr-12-04-5619: Effects of NLK on the cell cycle and cell apoptosis determined by flow cytometric analysis. (A) Histograms showing the cell cycle distribution of uninfected Hep-2 cells (Con) and cells infected with the Lv-shNLK and Lv-shCon. (B) Suppression of NLK expression by RNA interference resulted in an increase in the percentage of cells in the S phase. (C) A significant increase was identified in the percentage of cells in the sub-G1 phase following transfection with NLK siRNA. The percentage of cells in each phase of the cycle is presented as the mean ± standard error of three independent experiments. NLK, nemo-like kinase.

Mentions: The present study also aimed to examine the effects of NLK knockdown on cell cycle progression. Hep-2 cells were labeled with PI and analyzed by DNA flow cytometry. A representative histogram for the Hep-2 cells is shown in Fig. 3A. NLK knockdown caused an increase in the percentage of cells in the S phase and a corresponding decrease in the percentage of cells in the G0/G1 and G2-M phases (Fig. 3B). In addition, as depicted in Fig. 3C, the percentage of sub-G1 apoptotic cells of Lv-shNLK were significantly higher compared with Lv-shCon and Con.


Lentivirus‑delivered nemo‑like kinase small interfering RNA inhibits laryngeal cancer cell proliferation in vitro.

Tai J, Rao Y, Fang J, Huang Z, Yu Z, Chen X, Zhou W, Xiao X, Long T, Han Y, Liu Q, Li A, Ni X - Mol Med Rep (2015)

Effects of NLK on the cell cycle and cell apoptosis determined by flow cytometric analysis. (A) Histograms showing the cell cycle distribution of uninfected Hep-2 cells (Con) and cells infected with the Lv-shNLK and Lv-shCon. (B) Suppression of NLK expression by RNA interference resulted in an increase in the percentage of cells in the S phase. (C) A significant increase was identified in the percentage of cells in the sub-G1 phase following transfection with NLK siRNA. The percentage of cells in each phase of the cycle is presented as the mean ± standard error of three independent experiments. NLK, nemo-like kinase.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581764&req=5

f3-mmr-12-04-5619: Effects of NLK on the cell cycle and cell apoptosis determined by flow cytometric analysis. (A) Histograms showing the cell cycle distribution of uninfected Hep-2 cells (Con) and cells infected with the Lv-shNLK and Lv-shCon. (B) Suppression of NLK expression by RNA interference resulted in an increase in the percentage of cells in the S phase. (C) A significant increase was identified in the percentage of cells in the sub-G1 phase following transfection with NLK siRNA. The percentage of cells in each phase of the cycle is presented as the mean ± standard error of three independent experiments. NLK, nemo-like kinase.
Mentions: The present study also aimed to examine the effects of NLK knockdown on cell cycle progression. Hep-2 cells were labeled with PI and analyzed by DNA flow cytometry. A representative histogram for the Hep-2 cells is shown in Fig. 3A. NLK knockdown caused an increase in the percentage of cells in the S phase and a corresponding decrease in the percentage of cells in the G0/G1 and G2-M phases (Fig. 3B). In addition, as depicted in Fig. 3C, the percentage of sub-G1 apoptotic cells of Lv-shNLK were significantly higher compared with Lv-shCon and Con.

Bottom Line: The effects of NLK downregulation on Hep‑2 cell proliferation and cell cycle progression were analyzed using an MTT assay and flow cytometry, respectively.Downregulation of NLK also inhibited tumorigenesis and regulated the expression of cell cycle protein expression levels.Therefore, it was hypothesized that NLK is necessary for cell survival and tumorigenesis in laryngeal cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, Beijing 100045, P.R. China.

ABSTRACT
Laryngeal squamous cell carcinoma is the most common form of head and neck squamous cell carcinoma. Multiple approaches have been applied to treat this type of cancer; however, no significant improvement in survival rate has been achieved. In the present study, the role of nemo‑like kinase (NLK) in human laryngeal carcinoma Hep‑2 cells was investigated. NLK has been identified as an important regulator of cell growth, patterning and cell death in a variety of organisms. Lentivirus‑mediated‑shRNA was employed to silence endogenous NLK expression. Downregulation of the expression of NLK following lentivirus infection was confirmed using reverse transcription quantitative polymerase chain reaction and western blot analysis. The effects of NLK downregulation on Hep‑2 cell proliferation and cell cycle progression were analyzed using an MTT assay and flow cytometry, respectively. Downregulation of NLK also inhibited tumorigenesis and regulated the expression of cell cycle protein expression levels. Therefore, it was hypothesized that NLK is necessary for cell survival and tumorigenesis in laryngeal cancer cells. Furthermore, the absence of NLK may lead to cancer cell death. Collectively, the results of the present study demonstrated that the lentivirus‑mediated targeted disruption of NLK may be a promising therapeutic method for the treatment of laryngeal cancer.

Show MeSH
Related in: MedlinePlus