Tegillarca granosa Extract Haishengsu Induces Apoptosis in Human Hepatocellular Carcinoma Cell Line BEL-7402 Via Fas-Signaling Pathways

Abstract

This study was designed to investigate the apoptosis-inducing properties of Tegillarca granosa extract Haishengsu (HSS) in human hepatocellular carcinoma cell line BEL-7402. Proliferation inhibition of the human hepatocellular carcinoma BEL-7402 cells was determined by the MTT assay, and the cell viability was determined by trypan blue dye exclusion assay. Apoptosis of BEL-7402 cells was demonstrated by fluorescence microscope with flow cytometry with Annexin V-FITC/PI double staining and Hoechst 33258 staining. Western blot analysis and RT-PCR were used to determine the expression levels of Fas. Expressions of caspase-8 and caspase-3 were examined by caspase activity assay and western blot analysis. HSS inhibited the proliferation of human hepatocellular carcinoma BEL-7402 cells in a dose- and time-dependent manner. Our results showed HSS had positive effect on apoptosis through flow cytometry assay and fluorescence microscope. The expressions of Fas protein and mRNA were up-regulated following the treatment. Caspase-8 and caspase-3 were activated in the cells cultured with HSS. In conclusion, HSS induced apoptosis of human hepatocellular carcinoma BEL-7402 cells. The apoptosis was associated with the up-regulation of Fas and the activations of caspase-8 and caspase-3.

Tegillarca granosa extract Haishengsu inhibits tumor activity via a mitochondrial‑mediated apoptotic pathway

“…Our previous research demonstrated that HSS induces apoptosis of BEL-7402 cells, as determined by flow cytometric analysis and Hoechst 33258 nuclear staining (15). To further confirm these results, a DNA fragmentation assay was conducted, since DNA fragmentation is a classical hallmark of apoptosis.…”

“…Tegillarca granosa as described previously (15), and dissolved in phosphate-buffered saline (PBS) prior to use.…”

“…Furthermore, murine studies have indicated that HSS may inhibit the growth of Ehrlich ascites tumors and improve immunological functions (14). Our previous study demonstrated that HSS was able to induce the apoptosis of human hepatocellular carcinoma (HCC) cells in a concentration-dependent manner, through the Fas signaling pathway, with caspase-8 and caspase-3 activation (15).…”

“…Classical apoptosis can be initiated via two major pathways: The intrinsic or mitochondrial-mediated pathway, Tegillarca granosa extract Haishengsu inhibits tumor activity via a mitochondrial-mediated apoptotic pathway and the extrinsic or death receptor-mediated pathway (17). A study from our laboratory indicated that HSS induces apoptosis in HCCBEL-7402 cells via the Fas signaling pathway (15). However, little is currently known regarding the mechanisms responsible for the mitochondrial-mediated apoptotic pathway of HSS against HCC cells.…”

Marine drug Haishengsu increases chemosensitivity to conventional chemotherapy and improves quality of life in patients with acute leukemia

“…The natural products may have an adjuvant effect when added to conventional chemotherapy in patients with cancers [1,2]. Haishengsu (HSS) is a purified protein from Tegillarca granosa L. (Order Arcoida) which is a sea creature that contains a high concentration of proteins and vitamin B 12 [3][4][5][6][7][8][9][10][11]. When used as an adjuvant therapy, HSS was reported to improve clinical outcomes of patients undergoing conventional chemotherapy for lung or renal cancer [2,8].…”