Bryophyllum pinnatum (BP) has been used in anthroposophic medicine to treat “hyperactivity diseases” such as tocolytic symptom and overactive bladder syndrome since the beginning of 20th century with no side effect. Its major secondary metabolites include bufadienolides, flavonoids, triterpenes, and steroids. BP has been widely used in tropical regions as traditional medicine. It is reported to have antioxidant, immunomodulatory, antidiabetic, anti-inflammatory, antihypertensive, wound healing, cytotoxic, and antitumor promoting activities. Glioblastoma is an aggressive and lethal tumor of the brain with few treatment options. In this study, the effects of aqueous extracts of BP on the induction and modulation of apoptosis in glioblastoma U87-MG cells were assessed using the green/red/blue fluorescent Apoptosis/Necrosis Detection Kit and the Human Apoptosis Antibody Array - Membrane (43 Targets) test by the Abcam cooperation. Our data revealed that 3-hours of 1 mg/mL and 2 mg/mL BP treatment induced statistically similar amounts of apoptosis observed as green apoptotic cells and red necrotic cells in U87-MG cells respectively (apoptotic cells = 86.0 ± 6%, necrotic cells = 14.0 ± 6.0% ; apoptotic cells = 85.0 ± 4.0% , necrotic cells = 14.5 ± 3.5%). The percent of apoptosis was not significantly different as compared to that of the positive control induced by 1µMol Staurosporine (94.5 ± 0.5%, 5.5 ± 0.5%); while significantly different from the negative control which had none apoptotic cells. Slightly less induction of apoptosis was obtained from 6-hours of 1 mg/mL and 2 mg/mL BP treatment with more necrotic cells (81.5 ± 2.5%, 18.5 ± 2.5%; 78.0 ± 3.0%, 21.5 ± 2.5% respectively). Modulation of various apoptosis markers such as up-regulation of Bax, p53, Caspase 3; and down-regulation of Bcl-2 and Bcl-w was also observed. These results suggest that BP contains phytochemicals which induce apoptosis in glioblastoma U87-MG cells by modulating these apoptotic pathway markers. Further study of the specific modulation effects of BP on apoptosis is warranted to reveal its potential chemopreventive and therapeutic properties against glioblastoma and other cancers. Citation Format: Kristin Ferrer, Joshua Li, Camille Kordas, Jasmine Cha, Sung Been Han, Joon Seo, Nathaly Manrique, Min Seo Kang, Yi Shan Jin, Si Won Choi, Ben Hiramoto, Denise Smith, Brian Yuen Wong. Modulation of apoptosis in glioblastoma U87-MG cancer cells by aqueous extract of Bryophyllum pinnatum [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1923.
Scutellaria barbata (SB) and Oldenlandia diffusa (OD) have been used in traditional Chinese medicine for treating liver, lung and rectal tumors. These two herbs are included in most of the herbal cancer treatment formulas in Taiwan hospitals. We previously showed that aqueous extracts of SB and OD inhibited mutagenesis, DNA binding and metabolism of aflatoxin B1 and benzo(a)pyrene. They were also shown to inhibit foci formation in the colon of AOM-induced mice. Other researchers demonstrated their effects in regulation of various apoptotic enzymes and pathways. Ethanol extract of SB was shown to inhibit colorectal cancer growth by suppression of the Wnt/β-catenin signaling pathway while ethanol extract of OD was indicated in the inhibition of proliferation and induction of apoptosis of 5-FU resistant colorectal cancer cells via the regulation of PI3K/AKT signaling pathway. In this study, the effects of aqueous extracts of SB and OD on the induction and modulation of apoptosis in HCT 116 colon cancer cells and CCD 841 CoN normal color epithelial cells were assessed using TUNEL assay, Apoptosis/Necrosis Detection Kit, and Human Apoptosis Antibody Array - Membrane (43 Targets) test (Abcam). Our data demonstrated that both the 3-hours and 6 hours of 1.5 mg/mL SB treatments induced apoptosis in HCT 116 colon cancer cells significantly (77.5 ± 1.5%, 87.5± 3.5%) as compared to the negative control (13.5 ± 0.5%, 17.0 ± 6.0%), p < 0.05. The percent of apoptosis was not significantly different as compared to that of the positive control induced by 1µMol Staurosporine (99.5 ± 0.5%, 91.9 ± 9.0%); while there was no notable induction of apoptosis in normal CCD 841 CoN colon epithelial cells (18.5 ± 1.5%, 14.4 ± 4.0% separately). The majority of the normal cells remained non-induced (71.0 ± 1.0%, 68.5 ± 6.5%). A similar selective induction of apoptosis, in the two cell lines separately, was obtained from the 3-hours and 6 hours of 1.5 mg/mL OD treatments (HCT116 - 44.0 ± 4.0%, 83.5± 15.5%; CCD841 CoN - 6.5 ± 5.5%, 7.0 ± 2.0% respectively). Modulation of various apoptosis markers production such as up-regulation of Bax, BID, Bad, p27, p53, Caspases 8 and 3; and down-regulation of Bcl-2 and Bcl-w was also observed. These results suggested that SB and OD contain phytochemicals which selectively induce apoptosis in HCT 116 cancer cells by modulating these markers while not significantly affecting the normal CCD 841 CoN colon epithelial cells. Further study of their specific modulation effects and phytochemicals on apoptosis is warranted to reveal their potential chemopreventive and therapeutic properties against colon cancer and other cancers. Citation Format: Camille Kordas, Joshua Li, Kristin Ferrer, Yi Shan Jin, Nathaly Manrique, Min Seo Kang, Joon Seo, Sung Been Han, Jasmine Cha, Anthony Miller, Padma Tadi Uppala, Brian Yuen Wong. Selective induction of apoptosis by aqueous extract of Chinese medicinal herbs Scutellaria barbata and Oldenlandia diffusain HCT 116 colon cancer cells and CCD 841 CoN colon epithelial cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1926.
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