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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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Western blot analysis of apoptosis-associated proteins. Western blot analysis revealed that knockdown of NLK resulted in upregulation of pro-apoptotic proteins in Lv-shNLK cells, suggesting that the apoptotic effect of NLK may be partly mediated by pro-apoptotic family member proteins.
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f4-mmr-12-04-5619: Western blot analysis of apoptosis-associated proteins. Western blot analysis revealed that knockdown of NLK resulted in upregulation of pro-apoptotic proteins in Lv-shNLK cells, suggesting that the apoptotic effect of NLK may be partly mediated by pro-apoptotic family member proteins.

Mentions: In addition to the possible molecular mechanism for cell growth inhibition and cell cycle arrest in Hep-2 cells, pro-apoptotic protein expression was also investigated. The protein expression of cell cycle regulatory proteins, including cyclin B1, cyclin D1, p53 and pS315-p53 were examined (Fig. 4). Protein levels of cyclin D1, whose function is important for the G1-S transition, were significantly upregulated in Lv-shNLK. In addition, cyclin B1 expression, which controls G2-M transition was also upregulated in Lv-shNLK. These results demonstrated that NLK may be involved in translational modification of cell cycle regulatory proteins in Hep-2 cells. Furthermore, Lv-shNLK markedly phosphorylated p53 (Fig. 4), while uninfected cells and Lv-shCon did not exhibit phosphorylated p53. These results confirm that Lv-shNLK specifically phosphorylates p53 in vitro.


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)

Western blot analysis of apoptosis-associated proteins. Western blot analysis revealed that knockdown of NLK resulted in upregulation of pro-apoptotic proteins in Lv-shNLK cells, suggesting that the apoptotic effect of NLK may be partly mediated by pro-apoptotic family member proteins.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4-mmr-12-04-5619: Western blot analysis of apoptosis-associated proteins. Western blot analysis revealed that knockdown of NLK resulted in upregulation of pro-apoptotic proteins in Lv-shNLK cells, suggesting that the apoptotic effect of NLK may be partly mediated by pro-apoptotic family member proteins.
Mentions: In addition to the possible molecular mechanism for cell growth inhibition and cell cycle arrest in Hep-2 cells, pro-apoptotic protein expression was also investigated. The protein expression of cell cycle regulatory proteins, including cyclin B1, cyclin D1, p53 and pS315-p53 were examined (Fig. 4). Protein levels of cyclin D1, whose function is important for the G1-S transition, were significantly upregulated in Lv-shNLK. In addition, cyclin B1 expression, which controls G2-M transition was also upregulated in Lv-shNLK. These results demonstrated that NLK may be involved in translational modification of cell cycle regulatory proteins in Hep-2 cells. Furthermore, Lv-shNLK markedly phosphorylated p53 (Fig. 4), while uninfected cells and Lv-shCon did not exhibit phosphorylated p53. These results confirm that Lv-shNLK specifically phosphorylates p53 in vitro.

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