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Knockdown of GPC3 inhibits the proliferation of Huh7 hepatocellular carcinoma cells through down-regulation of YAP.
J Cell Biochem. 2013 Mar; 114(3):625-31.JC

Abstract

Glypican-3 (GPC3), a membrane-associated heparan sulfate proteoglycan, is frequently upregulated in hepatocellular carcinoma (HCC). Yes-associated protein (YAP) is also found over-expressed in HCC and has been identified as a key effector molecule in Hippo pathway, which could control the organ size in animals through the regulation of cell proliferation and apoptosis and plays an important role in the development of malignant tumors. Studies have reported that GPC3 and YAP might collaborate to regulate the development of HCC. To elucidate the role of GPC3 in the development of HCC and its relationship with YAP, siRNA technique was employed to knock down GPC3 in Huh7 HCC cells. Moreover, recombinant human YAP-1 was used to examine the effects of GPC3 on Huh7 cells. The results of flow cytometric analysis and Annexin-V-FLUOS apoptosis assay showed that knockdown of GPC3-induced apoptosis in Huh7 cells, resulting in inhibition of cell proliferation as examined by EdU incorporation assay, migration, and invasion. GPC3 knockdown also suppressed the expression of YAP in mRNA and protein levels, as examined by fluorescence quantitative PCR and Western blot analysis. Moreover, addition of recombinant human YAP-1 effectively rescued the cells from apoptosis triggered by GPC3 knockdown. Taken together, our findings suggest that GPC3 regulates HCC cell proliferation with the involvement of Hippo pathway.

Authors+Show Affiliations

Department of Hepatobiliary Surgery, Affiliated Hospital of Guangdong Medical College, Zhanjiang 524001, China. miaohuilai@tom.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23060277

Citation

Miao, Hui-Lai, et al. "Knockdown of GPC3 Inhibits the Proliferation of Huh7 Hepatocellular Carcinoma Cells Through Down-regulation of YAP." Journal of Cellular Biochemistry, vol. 114, no. 3, 2013, pp. 625-31.
Miao HL, Pan ZJ, Lei CJ, et al. Knockdown of GPC3 inhibits the proliferation of Huh7 hepatocellular carcinoma cells through down-regulation of YAP. J Cell Biochem. 2013;114(3):625-31.
Miao, H. L., Pan, Z. J., Lei, C. J., Wen, J. Y., Li, M. Y., Liu, Z. K., Qiu, Z. D., Lin, M. Z., Chen, N. P., & Chen, M. (2013). Knockdown of GPC3 inhibits the proliferation of Huh7 hepatocellular carcinoma cells through down-regulation of YAP. Journal of Cellular Biochemistry, 114(3), 625-31. https://doi.org/10.1002/jcb.24404
Miao HL, et al. Knockdown of GPC3 Inhibits the Proliferation of Huh7 Hepatocellular Carcinoma Cells Through Down-regulation of YAP. J Cell Biochem. 2013;114(3):625-31. PubMed PMID: 23060277.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Knockdown of GPC3 inhibits the proliferation of Huh7 hepatocellular carcinoma cells through down-regulation of YAP. AU - Miao,Hui-Lai, AU - Pan,Zhi-Jian, AU - Lei,Chang-Jiang, AU - Wen,Ji-Yu, AU - Li,Ming-Yi, AU - Liu,Zhong-Kao, AU - Qiu,Zhi-Dong, AU - Lin,Man-Zhou, AU - Chen,Nian-Ping, AU - Chen,Ming, PY - 2012/06/07/received PY - 2012/09/18/accepted PY - 2012/10/13/entrez PY - 2012/10/13/pubmed PY - 2013/7/10/medline SP - 625 EP - 31 JF - Journal of cellular biochemistry JO - J. Cell. Biochem. VL - 114 IS - 3 N2 - Glypican-3 (GPC3), a membrane-associated heparan sulfate proteoglycan, is frequently upregulated in hepatocellular carcinoma (HCC). Yes-associated protein (YAP) is also found over-expressed in HCC and has been identified as a key effector molecule in Hippo pathway, which could control the organ size in animals through the regulation of cell proliferation and apoptosis and plays an important role in the development of malignant tumors. Studies have reported that GPC3 and YAP might collaborate to regulate the development of HCC. To elucidate the role of GPC3 in the development of HCC and its relationship with YAP, siRNA technique was employed to knock down GPC3 in Huh7 HCC cells. Moreover, recombinant human YAP-1 was used to examine the effects of GPC3 on Huh7 cells. The results of flow cytometric analysis and Annexin-V-FLUOS apoptosis assay showed that knockdown of GPC3-induced apoptosis in Huh7 cells, resulting in inhibition of cell proliferation as examined by EdU incorporation assay, migration, and invasion. GPC3 knockdown also suppressed the expression of YAP in mRNA and protein levels, as examined by fluorescence quantitative PCR and Western blot analysis. Moreover, addition of recombinant human YAP-1 effectively rescued the cells from apoptosis triggered by GPC3 knockdown. Taken together, our findings suggest that GPC3 regulates HCC cell proliferation with the involvement of Hippo pathway. SN - 1097-4644 UR - https://www.unboundmedicine.com/medline/citation/23060277/Knockdown_of_GPC3_inhibits_the_proliferation_of_Huh7_hepatocellular_carcinoma_cells_through_down_regulation_of_YAP_ L2 - https://doi.org/10.1002/jcb.24404 DB - PRIME DP - Unbound Medicine ER -