Pinellia ternata extract induces apoptosis of leukemia cells by regulating the expression of Bax, Bcl-2 and Caspase-3 proteins

doi:10.3969/j.issn.2095-4344.2120

Abstract

Abstract:

BACKGROUND: Pinellia ternata can be cultivated artificially and is easy to purify. Its extract can inhibit the cell proliferation of chronic myeloid leukemia, acute myelogenous leukemia M3, and T-cell acute lymphoblastic leukemia, but the underlying mechanism remains unclear.

OBJECTIVE: To explore the mechanism of action of pinellia ternata extract on apoptosis of three kinds of leukemia cells.

METHODS: Chronic myeloid leukemia cell lines (K562), acute myeloid leukemia–M3 cell line (HL-60), acute T lymphocyte leukemia cell line (C8166) were incubated in five kinds of concentration in pinellia ternata extracts in vitro for 24, 48, and 72 hours, respectively. Cell counting kit-8 assay was used to assess the influence of three kinds of leukemia cell proliferation to screen the optimal inhibitory effect of moderate (300 mg/L) and high (500 mg/L) concentration. Flow cytometry was used to determine the incidence of early apoptosis of three kinds of leukemia cells. The mRNA and protein expression levels of Bax, Bcl-2 and Caspase-3 were detected by RT-PCR and western blot assay.

RESULTS AND CONCLUSION: Cell counting kit-8 assay results showed that five different concentrations of pinellia extracts could inhibit the proliferation of three kinds of leukemia cells. The inhibition rates of medium concentration (300 mg/L) and high concentration (500 mg/L) were significantly higher than those of low concentration (100 mg/L) and control group (P < 0.01). Flow cytometry (Annexin PE/7AAD and Annexin V/PI) double staining showed that moderate and high concentrations of pinellia extract could induce early apoptosis of three kinds of leukemia cells. RT-PCR results revealed moderate and high concentrations of pinellia extract could down-regulate the mRNA expression level of Bcl-2, and up-regulate the mRNA expression levels of Bax, Bcl-2 , Caspase-3 (P < 0.05, P < 0.01). Western blot assay results showed that the Bcl-2 protein bands of K562 and C8166 cells were significantly weakened after 72 hours of incubation with moderate and high concentrations of pinellia tubulin extract, while no significant changes were observed in HL-60 cells. Bax and caspase-3 protein bands were significantly enhanced. In summary, pinellia ternata extract can effectively inhibit the proliferation of myeloid leukemia and lymphocytic leukemia cells and promote their apoptosis via regulating Bax/Bcl-2 and Caspase-3 protein expression.

Key words: pinellia ternata extract, leukemia cells, apoptosis, Bax, Bcl-2, Caspase-3

CLC Number:

Feng Jiakun, Liu Wei, Li Zhengfa, Zhao Xiaochen, Li Zengzheng, Yang Tonghua, Zhao Renbin, Hu Peng, Pei Qiang, Guan Xin. Pinellia ternata extract induces apoptosis of leukemia cells by regulating the expression of Bax, Bcl-2 and Caspase-3 proteins[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(31): 5023-5029.

Figures/Tables 8

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