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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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Related in: MedlinePlus

NLK regulates the cell proliferation and colony formation of Hep-2 cells. (A) Cell proliferation in Hep-2 cells was measured using the MTT method as described in Materials and methods. Uninfected Hep-2 cells (Con) and cells infected with the Lv-shNLK and Lv-shCon were seeded in 6-well plates. (B) Colony formation assay. The number of colonies of Hep-2 cells following seeding for 14 days was counted. (C) Colonies stained with Giemsa were observed under fluorescence microscopy (magnification, ×200). (D) Hep-2 cells were seeded in 6 well plates at a density of 200 per well and were allowed to form colonies for 14 days. NLK, nemo-like kinase; OD, optical density.
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f2-mmr-12-04-5619: NLK regulates the cell proliferation and colony formation of Hep-2 cells. (A) Cell proliferation in Hep-2 cells was measured using the MTT method as described in Materials and methods. Uninfected Hep-2 cells (Con) and cells infected with the Lv-shNLK and Lv-shCon were seeded in 6-well plates. (B) Colony formation assay. The number of colonies of Hep-2 cells following seeding for 14 days was counted. (C) Colonies stained with Giemsa were observed under fluorescence microscopy (magnification, ×200). (D) Hep-2 cells were seeded in 6 well plates at a density of 200 per well and were allowed to form colonies for 14 days. NLK, nemo-like kinase; OD, optical density.

Mentions: To elucidate the role of NLK in cellular proliferation, the proliferative property of Lv-shNLK was analyzed using an MTT assay. The cell proliferation assay was performed for 5 days. The data revealed that there was a significant reduction in Hep-2 cell proliferation as a result of NLK knockdown compared with the uninfected cells (con) and Lv-shCon (Fig. 2A). Notably, 4 and 5 days post-infection, si-NLK-infected cells demonstrated a significantly lower cell viability of 0.58±0.00 and 0.60±0.01 fold, respectively, compared with the control group (P<0.01). These findings suggest that NLK is important in Hep-2 cell proliferation.


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)

NLK regulates the cell proliferation and colony formation of Hep-2 cells. (A) Cell proliferation in Hep-2 cells was measured using the MTT method as described in Materials and methods. Uninfected Hep-2 cells (Con) and cells infected with the Lv-shNLK and Lv-shCon were seeded in 6-well plates. (B) Colony formation assay. The number of colonies of Hep-2 cells following seeding for 14 days was counted. (C) Colonies stained with Giemsa were observed under fluorescence microscopy (magnification, ×200). (D) Hep-2 cells were seeded in 6 well plates at a density of 200 per well and were allowed to form colonies for 14 days. NLK, nemo-like kinase; OD, optical density.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2-mmr-12-04-5619: NLK regulates the cell proliferation and colony formation of Hep-2 cells. (A) Cell proliferation in Hep-2 cells was measured using the MTT method as described in Materials and methods. Uninfected Hep-2 cells (Con) and cells infected with the Lv-shNLK and Lv-shCon were seeded in 6-well plates. (B) Colony formation assay. The number of colonies of Hep-2 cells following seeding for 14 days was counted. (C) Colonies stained with Giemsa were observed under fluorescence microscopy (magnification, ×200). (D) Hep-2 cells were seeded in 6 well plates at a density of 200 per well and were allowed to form colonies for 14 days. NLK, nemo-like kinase; OD, optical density.
Mentions: To elucidate the role of NLK in cellular proliferation, the proliferative property of Lv-shNLK was analyzed using an MTT assay. The cell proliferation assay was performed for 5 days. The data revealed that there was a significant reduction in Hep-2 cell proliferation as a result of NLK knockdown compared with the uninfected cells (con) and Lv-shCon (Fig. 2A). Notably, 4 and 5 days post-infection, si-NLK-infected cells demonstrated a significantly lower cell viability of 0.58±0.00 and 0.60±0.01 fold, respectively, compared with the control group (P<0.01). These findings suggest that NLK is important in Hep-2 cell proliferation.

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