Trichosanthes kirilowii tuber extract induces G2/M phase arrest via inhibition of tubulin polymerization in HepG2 cells
Introduction
Many medical plants have served as anticancer pharmaceutical resources, and over 60% of current anticancer drugs such as vinblastine, topotecan, etoposide, and paclitaxel were originally plant-derived compounds (Newman et al., 2003, Cragg and Newman, 2005). Trichosanthes kirilowii tuber, an herbal plant of East Asia, traditionally has been prescribed for patients with diabetes, rigorous coughing, breast abscesses, and cancer-related symptoms. In the 1980s, a ribosome-inactivating protein, trichosanthin (TCS), was isolated from the tuber of this plant; it has been of interest because of its multiple pharmacological properties such as abortifacient, anti-HIV, immunoregulatory, and antitumor functions (Leung et al., 1986, Zheng et al., 1995, Zheng et al., 2000, Lu et al., 2001, Zhao et al., 2006). These TCS-derived pharmaceutical activities, including antitumor effects, were assumed to be connected to its biochemical activities, disrupting the binding of elongation factors to the P-complex as well as N-glycosidase activity for ribosome inactivation (Zhang and Liu, 1992, Chan et al., 2001, Shaw et al., 2005).
Thus far, many studies have focused on TCS, but seldom on other components of Trichosanthes kirilowii tuber as anticancer pharmaceutical resources. We screened numerous herbal plants and found that the methanol extract of Trichosanthes kirilowii tuber lacking TCS has interesting bioactivity related to the inhibition of cancer cell growth. In addition, a recent study demonstrated that the extract of Trichosanthes kirilowii tubers was superior to TCS in antitumor activity (Dou and Li, 2004).
We investigated the pharmaceutical effect of a 50% methanol extract of Trichosanthes kirilowii tubers on anti-cell growth and anti-tubulin polymerization causing M/G2 arrest in HepG2 cells.
Section snippets
Extraction of Trichosanthes kirilowii tuber and fingerprinting analysis
Dried and sliced Trichosanthes kirilowii tuber was provided from Dunsan Oriental Hospital of Daejeon University and identified by professor, SI Yim of Daejeon University. This herb (Voucher specimen number, TK-2006-01-Tu) was stocked at our laboratory for future reference. One hundred grams of powdered Trichosanthes kirilowii tuber was mixed with 1 L of 50% methanol and shaken for 48 h at 37 °C. The extract was filtered, dried out with a vacuum evaporator to remove methanol, redissolved with
Inhibition of cell proliferation without mortality
We first evaluated the antigrowth activity of TKE by determining the HepG2 cell growth rate during 48 h within media treated with various concentrations of TKE and 5-FU as a positive control. As shown in Fig. 2A, proliferation of HepG2 cells was significantly inhibited via treatment by TKE in a dose-dependent manner. The estimated IC50 of TKE in this cell line was approximately 25 μg/mL. Next, to determine whether these cells die or just stop growing from the treatment with TKE, we measured the
Discussion and conclusions
The tubers and fruits of Trichosanthes kirilowii have been a traditional herbal remedy and an attractive medical resource due to their novel pharmaceutical components (Oh et al., 2002, Fei et al., 2004, Kondo et al., 2004). Here, we found that Trichosanthes kirilowii methanol extract (TKE) induces cell cycle arrest at the G2/M phase and has activity inhibiting tubulin polymerization in HepG2 cells. This is the first study to report the pharmaceutical mechanisms of Trichosanthes kirilowii tubers
Acknowledgement
This study was supported by a grant from the Oriental Medicine R&D Project (B050018), Ministry of Health and Welfare, Republic of Korea.
References (30)
- et al.
Microtubule-interacting drugs for cancer treatment
Trends in Pharmacological Sciences
(2003) - et al.
Oncocidin A1: a novel tubulin-binding drug with antitumor activity against human breast and ovarian carcinoma xenografts in nude mice
Biochemical Pharmacology
(1998) - et al.
Plants as a source of anticancer agents
Journal of Ethnopharmacology
(2005) Curcumin disrupts mitotic spindle structure and induces micronucleation in MCF-7 breast cancer cells
Mutant Research
(2002)- et al.
The immunomodulatory and antitumor activities of trichosanthin—an abortifacient protein isolated from tian-hua-fen (Trichosanthes kirilowii)
Asian Pacific Journal of Allergy and Immunology
(1986) - et al.
The interaction of trichosanthin with supported phospholipid membranes studied by surface plasmon resonance
Biochimica et Biophysica Acta
(2001) - et al.
Effects of methyl mercury on the cell cycle of primary rat CNS cells in vitro
Toxicology and Applied Pharmacology
(1994) - et al.
Capillary electrophoretic determination of apoptosis of HeLa cells induced by trichosanthin
Journal of Chromatography A
(2000) - et al.
Recent advances in trichosanthin, a ribosome-inactivating protein with multiple pharmacological properties
Toxicon
(2005) - et al.
Cytotoxic synergy between flavopiridol (NSC 649890 L86-8275) and various antineoplastic agents: the importance of sequence of administration
Cancer Research
(1997)
Down-regulation of cyclin D1 by transcriptional repression in MCF-7 human breast cancer cells induced by flavopiridol
Cancer Research
Trichosanthin interacts with acidic ribosomal proteins P0 and P1 and mitotic checkpoint protein MAD2B
European Journal of Biochemistry
Effect of extracts of Trichosanthes root tubers on HepA-H cells and HeLa cells
World Journal of Gastroenterology
Isolation of a putative ribosome inactivating protein from dried roots of Trichosanthes kirilowii used in traditional Chinese medicine
Planta Medica
Abrogation of the Chk1 mediated G2 checkpoint pathway potentiates temozolomide-induced toxicity in a p53-independent manner in human glioblastoma cells
Cancer Research
Cited by (32)
The immune-enhancing effects of a mixture of Astragalus membranaceus (Fisch.) Bunge, Angelica gigas Nakai, and Trichosanthes Kirilowii (Maxim.) or its active constituent nodakenin
2022, Journal of EthnopharmacologyCitation Excerpt :Angelica gigas was reported to have various pharmacological properties including anti-bacterial, neuroprotective, anti-inflammatory, anti-oxidant, and anti-tumor properties in in vitro and in vivo models (Sowndhararajan and Kim, 2017). Trichosanthes Kirilowii has been traditionally used for diabetes, severe cough, and cancer-related symptoms (Shin et al., 2008). Trichosanthes Kirilowii was reported to have biological activities, such as anti-tumor and anti-HIV activities in in vitro models (Dat et al., 2010).
SH003 overcomes drug resistance and immune checkpoints by inhibiting JAK-STAT3 signaling in MCF7/ADR cells
2021, Phytomedicine PlusCitation Excerpt :SH003 is a mixture of three different herbs (Astragalus membranaceus, Angelica gigas, and Trichosanthes kirilowii). According to our previous report, A. membranaceus, A. gigas, and T. kirilowii showed anticancer effects in cell lines from several cancers, including leukemia, hepatocellular carcinoma, colon cancer, lung cancer, and gastric cancer (Cheng et al., 2004; Cho and Leung, 2007; Choi et al., 2014; Cui et al., 2003; Heo et al., 2009; Kongtun et al., 2009; Ku et al., 2018; Li et al., 2012; Shin et al., 2008). It was also reported that SH003 induces apoptosis, inhibits angiogenesis and suppresses the proliferation of tumor cells by inhibiting several pathways (Choi et al., 2016, 2014; Seo et al., 2017).
Lamprey immune protein-1 (LIP-1) from Lampetra japonica induces cell cycle arrest and cell death in HeLa cells
2018, Fish and Shellfish ImmunologyCitation Excerpt :Our study suggests that the overexpression of LIP-1 proteins is cytotoxic to HeLa cells. Cell death induced by the LIP-1 proteins occurred via the apoptotic pathway as a result of cell cycle arrest in the G2/M phase [18,19]. In the future, we will further identify the LIP-1 receptor molecules.
Albendazole as a promising molecule for tumor control
2016, Redox BiologyCitation Excerpt :Ours results (Fig. 8A and B) showed that ABZ besides inhibits G1 phase arrest G2/M phase in EAC carcinomas as expected, suggesting that ABZ mechanism of action in this cellular type appears to be related with tubulin polymerization inhibition [57]. In this context, microtubules (heterodimer composed of α tubulin and β tubulin subunits) are indispensable for the formation and disappearance of the mitotic spindle, in other words, they are responsible for separation of duplicated chromosomes during cell division, while disruption of microtubule dynamics by inhibiting polymerization or preventing depolymerization of tubulin, thereby promoting cell cycle arrest on G2/M phase [57,58]. MTX analysis showed that G1 phase was inhibited and S phase was arrested.
Multiple pathways were involved in tubeimoside-1-induced cytotoxicity of HeLa cells
2011, Journal of ProteomicsCitation Excerpt :Furthermore, Ma et al. published the similar findings in the recent study that TBMS1 increased the soluble fractions of tubulin in a time-dependent manner [43]. The reduction of polymerized tubulin was also shown to be critical for the G2/M cell cycle arrest induced by the extract of Trichosanthes kirilowii tuber [44]. Indeed, negative regulation of microtubule polymerization has been demonstrated to influence the cells entering into the cell cycle at mitosis [45].
Methanol Extract of Ficus pumila L. Inhibits Proliferation, Induces Apoptosis and Arrests the Cell Cycle in HepG2 Live Cancer Cells
2023, Asian Journal of Plant Sciences