Knockdown of GPC3 inhibits the proliferation of Huh7 hepatocellular carcinoma cells through down‐regulation of YAP

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.

Glypican-3 promotes epithelial-mesenchymal transition of hepatocellular carcinoma cells through ERK signaling pathway

“…Furthermore, we showed that both endogenous and exogenous GPC3 expression were correlated with metastasis and invasion. First, we found endogenous GPC3 expression was associate with HCC metastasis in HCC tissues and cells, which was consistent with the report from Miao et al that the knockdown of GPC3 resulted in the inhibition of cell migration and invasion in Huh7 cells (16). The migration and invasion capacities of HCC cell lines were also increased by exogenous GPC3 treatment, suggesting that in addition to endogenous GPC3, soluble/exogenous GPC3 may also function to promote HCC metastasis in patients.…”

“…Several studies have reported that GPC3 can stimulate the migration, and invasion (16,17) of HCC cell lines. Here, we focused on the clinical importance of GPC3 in HCC, and noted that there was a positive association between GPC3 expression and HCC metastasis progression.…”

“…Thus, GPC3 might be a more reliable tumor marker that could allow for an early diagnosis of HCC (14,15). In addition, several studies have shown that GPC3 promoted growth of HCC in vivo and in vitro (6,16,17). However, the clinical function of GPC3 in HCC progression remains obscure.…”

GPC3 reduces cell proliferation in renal carcinoma cell lines

“…Lin et al [19] and Murthy et al [40] found that GPC3 re-expression in ovarian cancer cell lines resulted in inhibition of the efficiency of colony formation. Some studies have shown that GPC3 plays an important role in cell growth and differentiation [41, 42], such as in the case of hepatocellular carcinoma [43]. The data obtained in the present study suggest that GPC3 inhibits cell proliferation in clear cell renal cell carcinoma.…”

Role of Hepatoprotectors and Immunomodulators in Regulation of Hepatocyte Apoptosis Induced by Antituberculosis Treatment

“…Участие некоторых гепатопротекторов и иммуномодуляторов в регуляции апоптоза гепатоцитов, индуцированного противотуберкулезными препаратами основного ряда Доказано, что апоптоз гепатоцитов играет важную роль в патогенезе многих заболеваний: вирусных гепати-тов [1,2], алкогольных поражений печени [3] и других па-тологических состояний [4,5]. В настоящее время отме-чено увеличение числа лекарственных поражений печени [6].…”