Polysaccharides Extracted from Rhizoma Pleionis Have Antitumor Properties In Vitro and in an H22 Mouse Hepatoma Ascites Model In Vivo

Abstract

Malignant ascites is a highly severe and intractable complication of advanced or recurrent malignant tumors that is often immunotherapy-resistant. Rhizoma Pleionis is widely used in traditional medicine as an antimicrobial and anticancer agent, but its effectiveness in treating malignant ascites is unclear. In the current study, we investigated the effect of polysaccharides isolated from Rhizoma Pleionis (PRP) on murine hepatocarcinoma H22 cells in an ascites model. We have found that the main components of PRP, that presented a relative molecular weight of 383.57 kDa, were mannose and glucose. We also found that PRP reduced the occurrence of abdominal ascites and increased survival in our mouse model. An immune response in the ascites tumor model was observed by performing a lymphocytes proliferation experiment and an E-rosette test. The ratios of CD8+ cytotoxic T cells and NK cells in the spleen were examined by flow cytometry, and the mRNA expression of Foxp3+in CD4+CD25+ (T regulatory Tregs) was measured by RT-PCR (reverse transcription-polymerase chain reaction). The levels of the cytokines TNF-α (tumor necrosis factor), VEGF (vascular endothelial growth factor), IL-2 (interleukin), and IFN-γ (interferon) in the serum and ascites supernatants were measured by ELISA. The expression of Foxp3 and Stat3 in peritoneal cells in the mouse model was measured by immunocytochemistry. The results indicated that PRP increased H22 tumor cell apoptosis in vivo by activating and enhancing the immune response. Furthermore, the effects of PRP on the proliferation of H22 cells were assessed by the CCK8 assay, Hoechest 33258, and TUNEL staining in vitro. We found that PRP suppressed the proliferation of H22 tumor cells but had no effect on BRL (Big rat liver) -3A rat hepatoma normal cells in vitro. Next, we investigated the underlying immunological mechanism by which PRP inhibits malignant ascites. PRP induced tumor cell apoptosis by inhibiting the Jak1–Stat3 pathway and by activating Caspase-3 and Caspase-8 to increase the Bax/Bcl-2 ratio. Collectively, our results indicate that PRP exhibits significant antitumor properties in H22 cells in vivo and in vitro, indicating that PRP may be used as a new therapeutic drug for cancer treatment.

Orchidaceae-Derived Anticancer Agents: A Review

“…The authors revealed that on the tenth day after tumor cell inoculation, the mouse abdominal perimeter and weight in the PRP treatment group were significantly smaller than those in the control group. Collectively, these results demonstrated that PRP has significant antitumour properties in the H22 tumor model [218]. Other studies in xenograft analysis showed that chrysotoxene (phenanthrene derivative that was first isolated from Dendrobium chrysotoxum) (20 mg/kg) indicated that it significantly (p < 0.01) the inhibited growth of HepG2 cell-induced tumors by regulating the aforementioned apoptotic proteins (Smac, Cytochrome c, Survivin, Bcl-2, Bax, Apaf-1, c-caspase-9, and c-caspase-3), compared with the control group.…”

Anti-tumor effect of polysaccharide from Pleurotus ostreatus on H22 mouse Hepatoma ascites in-vivo and hepatocellular carcinoma in-vitro model

“…2 that reflects the anti-ascites attributes of our extracted polysaccharide. Re-occurrence of tumor cells is one of the crucial hurdles in achieving successful cancer therapy (Fang et al 2018 ). Foxp3 and Stat3 genes were observed down-regulated in polysaccharide treated groups as compared to their control group that shows no signs of disease re-occurrence results presented in Fig.…”

Taxus wallichiana var. chinensis (Pilg.) Florin Aqueous Extract Suppresses the Proliferation and Metastasis in Lung Carcinoma via JAK/STAT3 Signaling Pathway

“…The lung cancer cell strain was examined as described previously ( Fang et al, 2018 ). Four types of human LC cell lines were measured by CCK-8 (Dojindo, Japan), as per the instruction.…”

“…Before the STAT3 expression level of two groups was detected by western blot, puromycin (puro) at the concentration of 2.5 μg/ ml was added for the selection of stable cell lines. The cell viabilities of stable strains were measured by CCK-8 (Dojindo, Japan) in the above method ( Fang et al, 2018 ), where the mediums were mixed with AETW at 0.5 mg/ml and only cultured for 48 h.…”