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Macrophage migration inhibitory factor as a potential prognostic factor in gastric cancer.
World J Gastroenterol. 2015 Sep 14; 21(34):9916-26.WJ

Abstract

AIM

To investigate macrophage migration inhibitory factor (MIF) expression and its clinical relevance in gastric cancer, and effects of MIF knockdown on proliferation of gastric cancer cells.

METHODS

Tissue microarray containing 117 samples of gastric cancer and adjacent non-cancer normal tissues was studied for MIF expression by immunohistochemistry (IHC) semiquantitatively, and the association of MIF expression with clinical parameters was analyzed. MIF expression in gastric cancer cell lines was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Two pairs of siRNA targeting the MIF gene (MIF si-1 and MIF si-2) and one pair of scrambled siRNA as a negative control (NC) were designed and chemically synthesized. All siRNAs were transiently transfected in AGS cells with Oligofectamine(TM) to knock down the MIF expression, with the NC group and mock group (Oligofectamine(TM) alone) as controls. At 24, 48, and 72 h after transfection, MIF mRNA was analyzed by RT-PCR, and MIF and proliferating cell nuclear antigen (PCNA) proteins were detected by Western blot. The proliferative rate of AGS cells was assessed by methylthiazolyl tetrazolium (MTT) assay and colony forming assay.

RESULTS

The tissue microarray was informative for IHC staining, in which the MIF expression in gastric cancer tissues was higher than that in adjacent non-cancer normal tissues (P < 0.001), and high level of MIF was related to poor tumor differentiation, advanced T stage, advanced tumor stage, lymph node metastasis, and poor patient survival (P < 0.05 for all). After siRNA transfection, MIF mRNA was measured by real-time PCR, and MIF protein and PCNA were assessed by Western blot analysis. We found that compared to the NC group and mock group, MIF expression was knocked down successfully in gastric cancer cells, and PCNA expression was downregulated with MIF knockdown as well. The cell counts and the doubling times were assayed by MTT 4 d after transfection, and colonies formed were assayed by colony forming assay 10 d after transfection; all these showed significant changes in gastric cancer cells transfected with specific siRNA compared with the control siRNA and mock groups (P < 0.001 for all).

CONCLUSION

MIF could be of prognostic value in gastric cancer and might be a potential target for small-molecule therapy.

Authors+Show Affiliations

Long-Jun He, Pin-Jin Hu, Sen-Lin Zhu, Department of Gastroenterology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China.Long-Jun He, Pin-Jin Hu, Sen-Lin Zhu, Department of Gastroenterology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China.Long-Jun He, Pin-Jin Hu, Sen-Lin Zhu, Department of Gastroenterology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China.Long-Jun He, Pin-Jin Hu, Sen-Lin Zhu, Department of Gastroenterology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China.Long-Jun He, Pin-Jin Hu, Sen-Lin Zhu, Department of Gastroenterology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China.Long-Jun He, Pin-Jin Hu, Sen-Lin Zhu, Department of Gastroenterology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China.Long-Jun He, Pin-Jin Hu, Sen-Lin Zhu, Department of Gastroenterology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

26379396

Citation

He, Long-Jun, et al. "Macrophage Migration Inhibitory Factor as a Potential Prognostic Factor in Gastric Cancer." World Journal of Gastroenterology, vol. 21, no. 34, 2015, pp. 9916-26.
He LJ, Xie D, Hu PJ, et al. Macrophage migration inhibitory factor as a potential prognostic factor in gastric cancer. World J Gastroenterol. 2015;21(34):9916-26.
He, L. J., Xie, D., Hu, P. J., Liao, Y. J., Deng, H. X., Kung, H. F., & Zhu, S. L. (2015). Macrophage migration inhibitory factor as a potential prognostic factor in gastric cancer. World Journal of Gastroenterology, 21(34), 9916-26. https://doi.org/10.3748/wjg.v21.i34.9916
He LJ, et al. Macrophage Migration Inhibitory Factor as a Potential Prognostic Factor in Gastric Cancer. World J Gastroenterol. 2015 Sep 14;21(34):9916-26. PubMed PMID: 26379396.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Macrophage migration inhibitory factor as a potential prognostic factor in gastric cancer. AU - He,Long-Jun, AU - Xie,Dan, AU - Hu,Pin-Jin, AU - Liao,Yi-Ji, AU - Deng,Hai-Xia, AU - Kung,Hsiang-Fu, AU - Zhu,Sen-Lin, PY - 2015/02/23/received PY - 2015/04/13/revised PY - 2015/07/15/accepted PY - 2015/9/18/entrez PY - 2015/9/18/pubmed PY - 2016/10/7/medline KW - Macrophage migration inhibitory factor KW - Proliferation KW - RNA interference KW - Stomach neoplasm KW - Survival SP - 9916 EP - 26 JF - World journal of gastroenterology JO - World J Gastroenterol VL - 21 IS - 34 N2 - AIM: To investigate macrophage migration inhibitory factor (MIF) expression and its clinical relevance in gastric cancer, and effects of MIF knockdown on proliferation of gastric cancer cells. METHODS: Tissue microarray containing 117 samples of gastric cancer and adjacent non-cancer normal tissues was studied for MIF expression by immunohistochemistry (IHC) semiquantitatively, and the association of MIF expression with clinical parameters was analyzed. MIF expression in gastric cancer cell lines was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Two pairs of siRNA targeting the MIF gene (MIF si-1 and MIF si-2) and one pair of scrambled siRNA as a negative control (NC) were designed and chemically synthesized. All siRNAs were transiently transfected in AGS cells with Oligofectamine(TM) to knock down the MIF expression, with the NC group and mock group (Oligofectamine(TM) alone) as controls. At 24, 48, and 72 h after transfection, MIF mRNA was analyzed by RT-PCR, and MIF and proliferating cell nuclear antigen (PCNA) proteins were detected by Western blot. The proliferative rate of AGS cells was assessed by methylthiazolyl tetrazolium (MTT) assay and colony forming assay. RESULTS: The tissue microarray was informative for IHC staining, in which the MIF expression in gastric cancer tissues was higher than that in adjacent non-cancer normal tissues (P < 0.001), and high level of MIF was related to poor tumor differentiation, advanced T stage, advanced tumor stage, lymph node metastasis, and poor patient survival (P < 0.05 for all). After siRNA transfection, MIF mRNA was measured by real-time PCR, and MIF protein and PCNA were assessed by Western blot analysis. We found that compared to the NC group and mock group, MIF expression was knocked down successfully in gastric cancer cells, and PCNA expression was downregulated with MIF knockdown as well. The cell counts and the doubling times were assayed by MTT 4 d after transfection, and colonies formed were assayed by colony forming assay 10 d after transfection; all these showed significant changes in gastric cancer cells transfected with specific siRNA compared with the control siRNA and mock groups (P < 0.001 for all). CONCLUSION: MIF could be of prognostic value in gastric cancer and might be a potential target for small-molecule therapy. SN - 2219-2840 UR - https://www.unboundmedicine.com/medline/citation/26379396/Macrophage_migration_inhibitory_factor_as_a_potential_prognostic_factor_in_gastric_cancer_ L2 - https://www.wjgnet.com/1007-9327/full/v21/i34/9916.htm DB - PRIME DP - Unbound Medicine ER -