Limits...
Role of ribophorin II in the response to anticancer drugs in gastric cancer cell lines.

Yuan TM, Liang RY, Chueh PJ, Chuang SM - Oncol Lett (2015)

Bottom Line: All the anticancer drugs were found to exert a concentration-dependent decrease on the cell survival rate of each of the cell lines tested, although the RPN2 levels in the various cell lines were not directly correlated with responsiveness to clinical anticancer drugs, based on the calculated IC50 values. siRNA-mediated RPN2 downregulation enhanced cisplatin-induced apoptosis in AGS cells, but did not markedly decrease the cell survival rates of these cells in response to the tested drugs.Furthermore, RPN2 silencing in MKN-45 cells resulted in no additional increase in the cisplatin-induced apoptosis and survival rates.However, the predictive value of RPN2 expression in cancer therapy is questionable in gastric cancer models.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biomedical Sciences, National Chung Hsing University, Taichung 40227, Taiwan, R.O.C. ; Department of Surgery, Feng-Yuan Hospital, Ministry of Health and Welfare, Taichung 42055, Taiwan, R.O.C.

ABSTRACT

The identification of prognostic markers and establishing their value as therapeutic targets improves therapeutic efficacy against human cancers. Ribophorin II (RPN2) has been demonstrated to be a prognostic marker of human cancer, including breast and pancreatic cancers. The present study aimed to evaluate RPN2 expression in gastric cancer and to examine the possible correlation between RPN2 expression and the response of cells to clinical anticancer drugs, which has received little research attention at present. The gastric cancer AGS, TMC-1, SNU-1, TMK-1, SCM-1, MKN-45 and KATO III cell lines were used as a model to elucidate the role of RPN2 in the response of cells to six common chemotherapeutic agents, comprising oxaliplatin, irinotecan, doxorubicin, docetaxel, cisplatin and 5-fluorouricil. The functional role of RPN2 was assessed by silencing RPN2 using small interfering RNA (siRNA), and the cytotoxicity was determined by an MTS assay and analysis of apoptosis. Molecular events were evaluated by western blotting. All the anticancer drugs were found to exert a concentration-dependent decrease on the cell survival rate of each of the cell lines tested, although the RPN2 levels in the various cell lines were not directly correlated with responsiveness to clinical anticancer drugs, based on the calculated IC50 values. siRNA-mediated RPN2 downregulation enhanced cisplatin-induced apoptosis in AGS cells, but did not markedly decrease the cell survival rates of these cells in response to the tested drugs. Furthermore, RPN2 silencing in MKN-45 cells resulted in no additional increase in the cisplatin-induced apoptosis and survival rates. It was also found that RPN2 depletion increased anticancer drug-mediated cytotoxicity in gastric cancer cell lines. However, the predictive value of RPN2 expression in cancer therapy is questionable in gastric cancer models.

No MeSH data available.


Related in: MedlinePlus

RPN2 silencing does not further increase cisplatin-induced cell death in the MKN-45 cell line. (A) The MKN-45 cells were transfected with siRPN2 to silence RPN2 expression. The protein levels of activated caspase-3, cleaved PARP, Bcl-2, RPN2, and β-actin in control and RPN2-knockdown cells following treatment with 2 μg/ml cisplatin were determined by western blot analysis. (B) The RPN2-knockdown MKN-45 cells were treated with 2.5 μg/ml cisplatin (IC50 for MKN-45) for 48 h, and the cell viability was determined using a MTS assay. The values were obtained from at least three independent experiments. PARP, poly(ADP-ribose) polymerase; Bcl-2, B-cell lymphoma 2; RPN2, ribophorin II; si, small interfering RNA; siC, control siRNA; siRPN2, RPN2 siRNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4356382&req=5

f6-ol-09-04-1861: RPN2 silencing does not further increase cisplatin-induced cell death in the MKN-45 cell line. (A) The MKN-45 cells were transfected with siRPN2 to silence RPN2 expression. The protein levels of activated caspase-3, cleaved PARP, Bcl-2, RPN2, and β-actin in control and RPN2-knockdown cells following treatment with 2 μg/ml cisplatin were determined by western blot analysis. (B) The RPN2-knockdown MKN-45 cells were treated with 2.5 μg/ml cisplatin (IC50 for MKN-45) for 48 h, and the cell viability was determined using a MTS assay. The values were obtained from at least three independent experiments. PARP, poly(ADP-ribose) polymerase; Bcl-2, B-cell lymphoma 2; RPN2, ribophorin II; si, small interfering RNA; siC, control siRNA; siRPN2, RPN2 siRNA.

Mentions: The functional significance of RPN2 in cell survival in response to six anticancer drugs was then investigated using MTS assays. The AGS cells were transfected with siRPN2 for 24 h and then treated with anticancer drugs for 48 h. In the presence of anticancer drugs, treatment with RPN2 siRNA slightly reduced the viability of AGS cells relative to the siRNA control. This effect of RPN2-knockdown was significant for all anticancer drugs with the exception of 5-FU, indicating that RPN2 may exert a protective role in cell survival (Fig. 5). RPN2 knockdown in MKN-45 cells also enhanced caspase 3 activation and Bcl-2 downregulation (Fig. 6A). However, subsequent treatment with cisplatin exhibited no evident effect on caspase 3 activation and demonstrated little synergetic effect on Bcl-2 downregulation. These observations were further supported by MTS assays, which revealed no significant decrease in survival in the cisplatin-exposed RPN2-knockdown MKN-45 cells compared with the cisplatin-exposed control siRNA cells (Fig. 6B).


Role of ribophorin II in the response to anticancer drugs in gastric cancer cell lines.

Yuan TM, Liang RY, Chueh PJ, Chuang SM - Oncol Lett (2015)

RPN2 silencing does not further increase cisplatin-induced cell death in the MKN-45 cell line. (A) The MKN-45 cells were transfected with siRPN2 to silence RPN2 expression. The protein levels of activated caspase-3, cleaved PARP, Bcl-2, RPN2, and β-actin in control and RPN2-knockdown cells following treatment with 2 μg/ml cisplatin were determined by western blot analysis. (B) The RPN2-knockdown MKN-45 cells were treated with 2.5 μg/ml cisplatin (IC50 for MKN-45) for 48 h, and the cell viability was determined using a MTS assay. The values were obtained from at least three independent experiments. PARP, poly(ADP-ribose) polymerase; Bcl-2, B-cell lymphoma 2; RPN2, ribophorin II; si, small interfering RNA; siC, control siRNA; siRPN2, RPN2 siRNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356382&req=5

f6-ol-09-04-1861: RPN2 silencing does not further increase cisplatin-induced cell death in the MKN-45 cell line. (A) The MKN-45 cells were transfected with siRPN2 to silence RPN2 expression. The protein levels of activated caspase-3, cleaved PARP, Bcl-2, RPN2, and β-actin in control and RPN2-knockdown cells following treatment with 2 μg/ml cisplatin were determined by western blot analysis. (B) The RPN2-knockdown MKN-45 cells were treated with 2.5 μg/ml cisplatin (IC50 for MKN-45) for 48 h, and the cell viability was determined using a MTS assay. The values were obtained from at least three independent experiments. PARP, poly(ADP-ribose) polymerase; Bcl-2, B-cell lymphoma 2; RPN2, ribophorin II; si, small interfering RNA; siC, control siRNA; siRPN2, RPN2 siRNA.
Mentions: The functional significance of RPN2 in cell survival in response to six anticancer drugs was then investigated using MTS assays. The AGS cells were transfected with siRPN2 for 24 h and then treated with anticancer drugs for 48 h. In the presence of anticancer drugs, treatment with RPN2 siRNA slightly reduced the viability of AGS cells relative to the siRNA control. This effect of RPN2-knockdown was significant for all anticancer drugs with the exception of 5-FU, indicating that RPN2 may exert a protective role in cell survival (Fig. 5). RPN2 knockdown in MKN-45 cells also enhanced caspase 3 activation and Bcl-2 downregulation (Fig. 6A). However, subsequent treatment with cisplatin exhibited no evident effect on caspase 3 activation and demonstrated little synergetic effect on Bcl-2 downregulation. These observations were further supported by MTS assays, which revealed no significant decrease in survival in the cisplatin-exposed RPN2-knockdown MKN-45 cells compared with the cisplatin-exposed control siRNA cells (Fig. 6B).

Bottom Line: All the anticancer drugs were found to exert a concentration-dependent decrease on the cell survival rate of each of the cell lines tested, although the RPN2 levels in the various cell lines were not directly correlated with responsiveness to clinical anticancer drugs, based on the calculated IC50 values. siRNA-mediated RPN2 downregulation enhanced cisplatin-induced apoptosis in AGS cells, but did not markedly decrease the cell survival rates of these cells in response to the tested drugs.Furthermore, RPN2 silencing in MKN-45 cells resulted in no additional increase in the cisplatin-induced apoptosis and survival rates.However, the predictive value of RPN2 expression in cancer therapy is questionable in gastric cancer models.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biomedical Sciences, National Chung Hsing University, Taichung 40227, Taiwan, R.O.C. ; Department of Surgery, Feng-Yuan Hospital, Ministry of Health and Welfare, Taichung 42055, Taiwan, R.O.C.

ABSTRACT

The identification of prognostic markers and establishing their value as therapeutic targets improves therapeutic efficacy against human cancers. Ribophorin II (RPN2) has been demonstrated to be a prognostic marker of human cancer, including breast and pancreatic cancers. The present study aimed to evaluate RPN2 expression in gastric cancer and to examine the possible correlation between RPN2 expression and the response of cells to clinical anticancer drugs, which has received little research attention at present. The gastric cancer AGS, TMC-1, SNU-1, TMK-1, SCM-1, MKN-45 and KATO III cell lines were used as a model to elucidate the role of RPN2 in the response of cells to six common chemotherapeutic agents, comprising oxaliplatin, irinotecan, doxorubicin, docetaxel, cisplatin and 5-fluorouricil. The functional role of RPN2 was assessed by silencing RPN2 using small interfering RNA (siRNA), and the cytotoxicity was determined by an MTS assay and analysis of apoptosis. Molecular events were evaluated by western blotting. All the anticancer drugs were found to exert a concentration-dependent decrease on the cell survival rate of each of the cell lines tested, although the RPN2 levels in the various cell lines were not directly correlated with responsiveness to clinical anticancer drugs, based on the calculated IC50 values. siRNA-mediated RPN2 downregulation enhanced cisplatin-induced apoptosis in AGS cells, but did not markedly decrease the cell survival rates of these cells in response to the tested drugs. Furthermore, RPN2 silencing in MKN-45 cells resulted in no additional increase in the cisplatin-induced apoptosis and survival rates. It was also found that RPN2 depletion increased anticancer drug-mediated cytotoxicity in gastric cancer cell lines. However, the predictive value of RPN2 expression in cancer therapy is questionable in gastric cancer models.

No MeSH data available.


Related in: MedlinePlus