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Role of Raf-kinase inhibitor protein in colorectal cancer and its regulation by hydroxycamptothecine.

Nie F, Cao J, Tong J, Zhu M, Gao Y, Ran Z - J. Biomed. Sci. (2015)

Bottom Line: Immunocytochemical analysis further revealed that RKIP expression was higher in the well differentiated cell line SW1116 as compared to that in the poorly differentiated cell line LoVo.Matrigel invasive assay demonstrated that the inhibition of RKIP by short hairpin RNA (shRNA) 271 transfection significantly increased the number of migrated cells (90.67 ± 4.04 vs. 37.33 ± 2.51, P <0.05), whereas over-expression of RKIP by PEBP-1 plasmid transfection significantly suppressed the number of migrated cells (79.24 ± 5.18 vs. 154.33 ± 7.25, P <0.05).Meanwhile, down-regulation of RKIP induced an increase in the cell survival rate by inhibiting apoptosis induced by hydroxycamptothecine.

View Article: PubMed Central - PubMed

Affiliation: Department of Intensive Care Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. niefang7@163.com.

ABSTRACT

Background: Recently accumulated evidence suggests that Raf kinase inhibitor protein (RKIP) participates in regulation of many signaling pathways and plays an important role in tumorigenesis and tumor metastasis. However, studies investigating the role of RKIP in colorectal cancer have not been reported. The aim of this study was to investigate the role of RKIP on colorectal cancer cell differentiation, progression and its correlation with chemosensitivity.

Results: Immunohistochemical analysis revealed that RKIP expression was higher in non-neoplastic colorectal tissue (NCRCT) and colorectal cancer tissue (CRCT) than that in metastatic lymph node tissue (MLNT) (P <0.05). P-ERK protein expression was higher in MLNT and CRCT than that in NCRCT (P = 0.02). Immunocytochemical analysis further revealed that RKIP expression was higher in the well differentiated cell line SW1116 as compared to that in the poorly differentiated cell line LoVo. Matrigel invasive assay demonstrated that the inhibition of RKIP by short hairpin RNA (shRNA) 271 transfection significantly increased the number of migrated cells (90.67 ± 4.04 vs. 37.33 ± 2.51, P <0.05), whereas over-expression of RKIP by PEBP-1 plasmid transfection significantly suppressed the number of migrated cells (79.24 ± 5.18 vs. 154.33 ± 7.25, P <0.05). Meanwhile, down-regulation of RKIP induced an increase in the cell survival rate by inhibiting apoptosis induced by hydroxycamptothecine.

Conclusions: RKIP was also found to be associated with cell differentiation, with a higher activity in well differentiated colorectal cancer cells than in poorly differentiated ones. The upregulated expression of RKIP in colorectal cancer cells inhibited cell invasion and metastasis, while downregulation of RKIP reduced chemosensitivity by inhibiting apoptosis induced by HCPT.

No MeSH data available.


Related in: MedlinePlus

The role of RKIP gene on cell cycle arrest and cell proliferation; cell cycle arrest and cell proliferation index as analyzed by flow cytometry: Down-regulation of RKIP inhibited G1 cell cycle arrest and promoted cell proliferation. G0/G1 %, G2/M % and PI % are significantly higher in SW1116/RKIP— cells with RKIP down-regulation, as compared to control SW1116 cells; Abbreviations: RKIP, Raf kinase inhibitor protein; PI %, Proliferation index
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Fig6: The role of RKIP gene on cell cycle arrest and cell proliferation; cell cycle arrest and cell proliferation index as analyzed by flow cytometry: Down-regulation of RKIP inhibited G1 cell cycle arrest and promoted cell proliferation. G0/G1 %, G2/M % and PI % are significantly higher in SW1116/RKIP— cells with RKIP down-regulation, as compared to control SW1116 cells; Abbreviations: RKIP, Raf kinase inhibitor protein; PI %, Proliferation index

Mentions: To avoid the impact of transfection reagent on cell cycle, we selected SW1116/RKIP— cells with steady down-regulation of RKIP expression, instead of SW1116 cells transfected with RKIP shRNA. The percentage of cells at S and G2/M phases by FCM was analysed by calculating the PI. PI % = (S + G2/M)/(G0/G1 + S + G2/M)×100 was calculated. Down-regulation of RKIP decreased the cells in G0/G1 phase (48.6 ± 2.12 vs. 57.6 ± 3.35, P <0.05) and increased the cells in G2/M phase (23.3 ± 4.31 vs. 14.9 ± 2.35, P <0.05), thereby increasing the PI (51.4 ± 2.12 vs. 42.4 ± 3.35, P <0.05) (Fig. 6). This suggests an important effect of RKIP on the invasion and migration of SW1116 and LoVo cells, and that down-regulation of RKIP inhibits G1 cell cycle arrest and promotes cell proliferation. At the same time, LoVo cells transfected with pIRES2-EGFP/PEBP-1 and LoVo cells transfected with pIRES2-EGFP negative control plasmid were analyzed through flow cytometry to observe the effect of over-expression of RKIP gene on the ability of cell cycle. There were no significant difference between the two groups.Fig. 6


Role of Raf-kinase inhibitor protein in colorectal cancer and its regulation by hydroxycamptothecine.

Nie F, Cao J, Tong J, Zhu M, Gao Y, Ran Z - J. Biomed. Sci. (2015)

The role of RKIP gene on cell cycle arrest and cell proliferation; cell cycle arrest and cell proliferation index as analyzed by flow cytometry: Down-regulation of RKIP inhibited G1 cell cycle arrest and promoted cell proliferation. G0/G1 %, G2/M % and PI % are significantly higher in SW1116/RKIP— cells with RKIP down-regulation, as compared to control SW1116 cells; Abbreviations: RKIP, Raf kinase inhibitor protein; PI %, Proliferation index
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig6: The role of RKIP gene on cell cycle arrest and cell proliferation; cell cycle arrest and cell proliferation index as analyzed by flow cytometry: Down-regulation of RKIP inhibited G1 cell cycle arrest and promoted cell proliferation. G0/G1 %, G2/M % and PI % are significantly higher in SW1116/RKIP— cells with RKIP down-regulation, as compared to control SW1116 cells; Abbreviations: RKIP, Raf kinase inhibitor protein; PI %, Proliferation index
Mentions: To avoid the impact of transfection reagent on cell cycle, we selected SW1116/RKIP— cells with steady down-regulation of RKIP expression, instead of SW1116 cells transfected with RKIP shRNA. The percentage of cells at S and G2/M phases by FCM was analysed by calculating the PI. PI % = (S + G2/M)/(G0/G1 + S + G2/M)×100 was calculated. Down-regulation of RKIP decreased the cells in G0/G1 phase (48.6 ± 2.12 vs. 57.6 ± 3.35, P <0.05) and increased the cells in G2/M phase (23.3 ± 4.31 vs. 14.9 ± 2.35, P <0.05), thereby increasing the PI (51.4 ± 2.12 vs. 42.4 ± 3.35, P <0.05) (Fig. 6). This suggests an important effect of RKIP on the invasion and migration of SW1116 and LoVo cells, and that down-regulation of RKIP inhibits G1 cell cycle arrest and promotes cell proliferation. At the same time, LoVo cells transfected with pIRES2-EGFP/PEBP-1 and LoVo cells transfected with pIRES2-EGFP negative control plasmid were analyzed through flow cytometry to observe the effect of over-expression of RKIP gene on the ability of cell cycle. There were no significant difference between the two groups.Fig. 6

Bottom Line: Immunocytochemical analysis further revealed that RKIP expression was higher in the well differentiated cell line SW1116 as compared to that in the poorly differentiated cell line LoVo.Matrigel invasive assay demonstrated that the inhibition of RKIP by short hairpin RNA (shRNA) 271 transfection significantly increased the number of migrated cells (90.67 ± 4.04 vs. 37.33 ± 2.51, P <0.05), whereas over-expression of RKIP by PEBP-1 plasmid transfection significantly suppressed the number of migrated cells (79.24 ± 5.18 vs. 154.33 ± 7.25, P <0.05).Meanwhile, down-regulation of RKIP induced an increase in the cell survival rate by inhibiting apoptosis induced by hydroxycamptothecine.

View Article: PubMed Central - PubMed

Affiliation: Department of Intensive Care Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. niefang7@163.com.

ABSTRACT

Background: Recently accumulated evidence suggests that Raf kinase inhibitor protein (RKIP) participates in regulation of many signaling pathways and plays an important role in tumorigenesis and tumor metastasis. However, studies investigating the role of RKIP in colorectal cancer have not been reported. The aim of this study was to investigate the role of RKIP on colorectal cancer cell differentiation, progression and its correlation with chemosensitivity.

Results: Immunohistochemical analysis revealed that RKIP expression was higher in non-neoplastic colorectal tissue (NCRCT) and colorectal cancer tissue (CRCT) than that in metastatic lymph node tissue (MLNT) (P <0.05). P-ERK protein expression was higher in MLNT and CRCT than that in NCRCT (P = 0.02). Immunocytochemical analysis further revealed that RKIP expression was higher in the well differentiated cell line SW1116 as compared to that in the poorly differentiated cell line LoVo. Matrigel invasive assay demonstrated that the inhibition of RKIP by short hairpin RNA (shRNA) 271 transfection significantly increased the number of migrated cells (90.67 ± 4.04 vs. 37.33 ± 2.51, P <0.05), whereas over-expression of RKIP by PEBP-1 plasmid transfection significantly suppressed the number of migrated cells (79.24 ± 5.18 vs. 154.33 ± 7.25, P <0.05). Meanwhile, down-regulation of RKIP induced an increase in the cell survival rate by inhibiting apoptosis induced by hydroxycamptothecine.

Conclusions: RKIP was also found to be associated with cell differentiation, with a higher activity in well differentiated colorectal cancer cells than in poorly differentiated ones. The upregulated expression of RKIP in colorectal cancer cells inhibited cell invasion and metastasis, while downregulation of RKIP reduced chemosensitivity by inhibiting apoptosis induced by HCPT.

No MeSH data available.


Related in: MedlinePlus