• Title/Summary/Keyword: Hep G2 cell

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Enterocarpam-III Induces Human Liver and Breast Cancer Cell Apoptosis via Mitochondrial and Caspase-9 Activation

  • Banjerdpongchai, Ratana;Wudtiwai, Benjawan;Pompimon, Wilart
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.1833-1837
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    • 2015
  • An aristolactam-type alkaloid, isolated from Orophea enterocarpa, is enterocarpam-III (10-amino-2,3,4,6-tetramethoxyphenanthrene-1-carboxylic acid lactam). It is cytotoxic to various human and murine cancer cell lines; however, the molecular mechanisms remain unclear. The aims of this study were to investigate cytotoxic effects on and mechanism (s) of human cancer cell death in human hepatocellular carcinoma HepG2 and human invasive breast cancer MDA-MB-231 cells compared to normal murine fibroblast NIH3T3 cells. Cell viability was determined by MTT assay to determine $IC_{10}$, $IC_{20}$ and $IC_{50}$ levels, reactive oxygen species (ROS) production with 2',7'-dichlorohydrofluorescein diacetate and the caspase-3, -8 and -9 activities using specific chromogenic (p-nitroaniline) tetrapeptide substrates, viz., DEVD-NA, IETD-NA and LEHD-NA and employing a microplate reader. Mitochondrial transmembrane potential (MTP) was measured by staining with 3, 3'-dihexyloxacarbocyanine iodide ($DiOC_6$) and using flow cytometry. The compound was cytotoxic to HepG2 and MDA-MB-231 cells with the $IC_{50}$ levels of $26.0{\pm}4.45$ and $51.3{\pm}2.05{\mu}M$, respectively. For murine normal fibroblast NIH3T3 cells, the $IC_{50}$ concentration was $81.3{\pm}10.1{\mu}M$. ROS production was reduced in a dose-response manner in HepG2 cells. The caspase-9 and -3 activities increased in a concentration-dependent manner, whereas caspase-8 activity did not alter, indicating the intrinsic pathway activation. Enterocarpam-III decreased the mitochondrial transmembrane potential (MTP) dose-dependently in HepG2 cells, suggesting that the compound induced HepG2 cell apoptosis via the mitochondrial pathway. In conclusion, enterocarpam-III inhibited HepG2 and MDA-MB-231 cell proliferation and induced human HepG2 cells to undergo apoptosis via the intrinsic (mitochondrial) pathway and induction of caspase-9 activity.

An Analytical Research of ONGABO with the Sovereign, Minister, Assistant and Courier Principle (君臣佐使論) on the Basis of HepG2 Cell Viability (간암세포주의 세포생존율에 근거한 온가보(溫家寶)(홍삼, 당귀, 오미자, 토사자, 울금)의 군신좌사론적(君臣佐使論的) 해석(解釋))

  • Hwang, Sung-Yeoun;Jeong, Kyung-Chae;Ahn, Seong-Hun
    • Journal of TMJ Balancing Medicine
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    • v.3 no.1
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    • pp.15-22
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    • 2013
  • Objectives and Methods: This study was conducted to investigate the formula of ONGABO to composed of Ginseng Radix (Red Ginseng), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen, Curcumae Tuber with the method to observe the cell viability of HepG2 in the basic principle of oriental medicine formula study, Sovereign, Minister, Assistant and Courier principle (君臣佐使論). Results: Ginseng Radix (Red Ginseng) and Schisandrae Fructus were having a cell protection effect in HepG2 significantly. Angelica gigantis radix was decreased the cell viability of HepG2 significantly, and there were no effects for Cuscuta Semen and Curcumae Tuber to the cell viability of HepG2. Conclusions: As the above results, in the Sovereign, Minister, Assistant and Courier principle (君臣佐使論), Ginseng Radix (Red Ginseng) corresponds to sovereign medicinal having cell protect effects, angelica gigantis radix corresponds to minister medicinal having cell killing effects, Schisandrae Fructus corresponds to assistant medicinal to help red ginseng having cell protect effects. Cuscuta Semen and Curcumae Tuber correspond to courier medicinal having no effect in cell viability in HepG2. We hope the advanced research on sovereign, minister, assistant and courier principle will be proceed in the tomorrow.

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Anti-proliferative Effect of Coptis Chinensis Extract in Hep G2 Cells

  • Kim, Jun-Lae;Oh, Se-Mi;Shin, Jang-Woo;Son, Jin-Young;Cho, Jung-Hyo;Lee, Yeon-Weol;Son, Chang-Gue;Cho, Chong-Kwan;Yoo, Hwa-Seung
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.48-56
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    • 2006
  • Objectives : This study is aimed to elucidate anti-hepatoma activity of Coptis Chinensis Extract (CCE) and evaluate its effect on proliferation of human hepatoma Hep G2 cells. Methods : To identify CCE and control the quality, we performed fingerprinting by high-performance thin layer chromatography (HPTLC). To investigate effects of CCE on anti-hepatoma activity, we measured cytotoxicity against Hep G2 cells compared with treatment of paclitaxel and 5-fluorouracil (5-FU). To examine the mechanism of inhibitory effect of CCE on Hep G2 cell proliferation, cell cycle distribution was evaluated using fluorescent activated cell sorter (FACS) Result : CCE showed a significant effect that arrests Hep G2 cells at the G2/M phase of the cell cycle. CCE combined with paclitaxel inhibited synergistically cell growth of Hep G2 cells. Conclusion : CCE may present anticancer effects through inhibition of hepatocellular carcinoma (HCC) cell proliferation via G2/M arrest, and may be a useful anticancer agent for HCC.

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Cytotoxic and Apoptotic-inducing Effects of Purple Rice Extracts and Chemotherapeutic Drugs on Human Cancer Cell Lines

  • Banjerdpongchai, Ratana;Wudtiwai, Benjawan;Sringarm, Korawan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6541-6548
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    • 2013
  • Pigmented rice is mainly black, red, and dark purple, and contains a variety of flavones, tannin, polyphenols, sterols, tocopherols, ${\gamma}$-oryzanols, amino acids, and essential oils. The present study evaluated the cytotoxic effects of purple rice extracts (PREs) combined with chemotherapeutic drugs on human cancer cells and mechanisms of cell death. Methanolic (MeOH) and dichloromethane (DCM) extracts of three cultivars of purple rice in Thailand: Doisaket (DSK), Nan and Payao (PYO), were tested and compared with white rice (KK6). Cytotoxicity was determined by 3-(4, 5-dimethyl)-2, 5-diphenyltetrazolium bromide (MTT) assay in human hepatocellular carcinoma HepG2, prostate cancer LNCaP and murine normal fibroblast NIH3T3 cells. MeOH-PYO-PRE was the most cytotoxic and inhibited HepG2 cell growth more than that of LNCaP cells but was not toxic to NIH3T3 cells. When PREs were combined with paclitaxel or vinblastine, they showed additive cytotoxic effects on HepG2 and LNCaP cells, except for MeOH-PYO-PRE which showed synergistic effects on HepG2 cells when combined with vinblastine. MeOH-PYO-PRE plus vinblastine induced HepG2 cell apoptosis with loss of mitochondrial transmembrane potential (MTP) but no ROS production. MeOH-PYO-PRE-treated HepG2 cells underwent apoptosis via caspase-9 and-3 activation. The level of ${\gamma}$-oryzanol was highest in DCM-PYO-PRE (44.17 mg/g) whereas anthocyanin content was high in MeOH-PYO-PRE (5.80 mg/g). In conclusion, methanolic Payao purple rice extract was mostly toxic to human HepG2 cells and synergistically enhanced the cytotoxicity of vinblastine. Human HepG2 cell apoptosis induced by MeOH-PYO-PRE and vinblastine was mediated through a mitochondrial pathway.

Studies of the Anti-cancer Effects of Bistortae Rhizoma (권삼(拳蔘)의 항암효과에 대한 연구)

  • Kim, June-Beom;Han, Hyo-Sang;Lee, Young-Jong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.5
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    • pp.1139-1144
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    • 2009
  • This study was purposed to research the anti-cancer effects of Bistortae Rhizoma. A total extract of Bistortae Rhizoma decoction was prepared. By measuring the cell proliferation, apoptosis, morphology and cytokine level from the extracts, the influence on HepG2 cell, SNU-1 cell and A549 cell was compared. The Bistortae Rhizoma decoction extract did not control HepG2 cell proliferation but controlled SNU-1 cell and A549 cell proliferation. In particular, the inhibitory effect on SNU-1 cell proliferation was highest. The Bistortae Rhizoma decoction extract showed to increase the apoptosis of the HepG2 ceil, SNU-1 cell and A549 cell in a dose-dependent manner. In particular, the promotion effect of the apoptosis was highest in SNU-1 cell. Among the various fraction extracts of the Bistortae Rhizoma decoction, n-BuOH extraction showed the greatest increase of the apoptosis of the HepG2 cell. The Bistortae Rhizoma decoction extract decreased dose-dependently the secretion of the TGF-$\beta$ in the HepG2 cell, SNU-1 cell and A549 cell and increased the secretion of the TNF-$\alpha$ and the IFN-$\gamma$. These results suggest that the total extract of Bistortae Rhizoma decoction has anti-cancer effect against SNU-1 cell and A549 cell.

The Combined Effects of n-BuOH Fraction of Ulmi Cortex and Anticancer Drugs on Cancer Cell Lines (암세포주에 대한 유근피 n-BuOH 분획과 항암제의 병용효과)

  • Eun, Jae-Soon;Song, Won-Young
    • Korean Journal of Pharmacognosy
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    • v.25 no.2
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    • pp.144-152
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    • 1994
  • The combined effects of Ulmi Cortex and some anti-cancer drugs on the proliferation of HeLa cells, Hep G2 cells and S 180 cells were estimated by MTT calorimetric assay. The n-BuOH fraction(UBF) of Ulmi Cortex inhibited the proliferation of HeLa cell at $10^{-3}\;g/ml$, Hep G2 cell at $10^{-5}\;g/ml$ and S 180 cell at $10^{-3}\;g/ml$. The inhibitory effects of mitomycin C(MMC), cisplatin(CPT) and 5-fluorouracil (5-FU), respectively, on Hep G2 cell was increased by the UBF. The UBF did not influence the proliferation of Balb/c 3T3 cells at concentrations of $10^{-6}$ to $10^{-4}\;g/ml$, but increased the proliferation of T cells at concentrations of $10^{-5}$ to $10^{-4}\;g/ml$. The UBF did not influence the number of leukocyte, and on the thymus weight of mice. The UBF increased the number of total-peritoreal cells of mice. In conclusion, the results suggest that the UBF have anti-cancer activity without the side effect, such as leukopenia and immunosuppresion, and increase the inhibitory activity of the anti-cancer drugs on Hep G2 cells.

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Effect of the Ethanol Extract of Vitis labrusca Root on Apoptosis in Hep G2 Cells (포도근 에탄올 추출물이 Hep G2 세포의 자연사에 미치는 효과)

  • Lee, Dong-Kyo;Lee, Kang-Pa;Kim, Hyuck;Choi, Byung-Jin;Chang, Hae-Ryong;Park, Won-Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.2
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    • pp.377-384
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    • 2008
  • The root of Vitis labrusca, is used as a source of health promoting drug in Korean traditional medicine. It has been reported that root of Vitis labrusca has antioxidant, anti lipid peroxidation and anti-reactive nitrogen species (RNS) activities. The aim of this study was to elucidate the molecular changes of apoptotic signaling pathways in phorbol 12-myristate 13 acetate (PMA)-induced human hepatocellular carcinoma cell line (Hep G2). The root of Vitis labrusca, ethanol extract (RVLEE) was tested for cell viability on Hep G2 cell using the MTT assay. RVLEE exhibited weak cytotoxic activity. However, treatment of Hep G2 cells with RVLEE suppressed PMA-induced cell proliferation. Also, dramatic changes of cell death signals in cellular molecules such as Chk2/Cds1, CIDE-B, CLIMP-63, Bax, Bcl-xL, C-myc, Bcl-2, Bric-5, NIP-3, TRAF2 and BAR but not CIDE-B and DR4. Futhermore, our results showed that the treatment of Hep G2 cells with 25 and $50\; {\mu}g/ml$ of RVLEE suppressed PMA-induced COX-2 gene activity. These data suggest that RVLEE have inhibitory effect of cell proliferation, induction of apoptosis and, thus, may offer therapeutic potential in Hep G2.

mRNA Expression of Bax, Bcl-2, p53, Cathepsin B, Caspase-3 and Caspase-9 in the HepG2 Cell Line Following Induction by a Novel Monoclonal Ab Hep88 mAb: Cross-Talk for Paraptosis and Apoptosis

  • Mitupatum, Thantip;Aree, Kalaya;Kittisenachai, Suthathip;Roytrakul, Sittiruk;Puthong, Songchan;Kangsadalampai, Sasichai;Rojpibulstit, Panadda
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.703-712
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    • 2016
  • Monoclonal antibodies with specific antigens have been widely used as targeted therapy for cancer. Hep88 mAb is a monoclonal antibody which shows specific binding with anti-cancer effects against the HepG2 cell line. However, its mechanisms of action are still not completely understood. We examined cell cycling and apoptosis by flow cytometry and mRNA expression of factors involved in apoptosis and paraptosis in Hep88 mAb-treated HepG2 cells by real-time PCR. The cell-cycle analysis demonstrated that growth-inhibitory activity was associated with G2/M cell cycle arrest. Hep88 mAb induced a significant increase in apoptotic cell populations in a dose- and time-dependent manner. The mRNA expression results also suggested that the process triggered by Hep88 mAb involved up-regulation of tumor suppressor p53, pro-apoptotic Bax, Cathepsin B, Caspase-3 and Caspase-9, with a decrease of anti-apoptotic Bcl-2 - thus confirming paraptosis and apoptosis programmed cell death. These findings represent new insights into the molecular mechanisms underlying the anti-cancer properties of Hep88 mAb in liver cancer cells.

Analytical Research to Determine the effects of the Components of ONGABO on the Viability of HepG2 Cancer Cells by Using the Sovereign, Minister, Assistant and Courier Principle (君臣佐使論)

  • Shin, Jeong-Hun;Jun, Seung-Lyul;Hwang, Sung-Yeoun;Ahn, Seong-Hun
    • Journal of Pharmacopuncture
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    • v.15 no.4
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    • pp.42-51
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    • 2012
  • Objectives: This study used the basic principle of Oriental medicine, the sovereign, minister, assistant and courier principle (君臣佐使論) to investigate the effects of the component of ONGABO, which is composed of Ginseng Radix (Red Ginseng), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen and Curcumae tuber on the viability of HepG2 cells. Methods: Single and mixed extracts of the component of ONGABO were prepared by lypohilizing powder of Red Ginseng (6-year root from Kanghwa), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen, Curcumae Tuber (from Omniherb Co., Ltd., Korea) at the laboratory of herbal medicine in Woosuk University and were eluted after being macerated with 100% ethanol for three days. The cell viability of HepG2 was determined by using an absorptiometric analysis with PrestoBlue (Invitrogen) reagent after the plate had been incubated for 48 hours. All of the experiments were repeated three times to obtain the average value and standard deviation. The statistical analysis was done and the correlation factor was obtained by using Microsoft Office Excel 2007 and Origin 6.0 software. Results: Although Ginseng Radix (Red Ginseng) and Schisandrae Fructus did not enhance the viability of HepG2 cells, they were shown to provide protection of those cells. On the other hand, Angelica Gigantis Radix decreased the viability of HepG2 cells significantly, Cuscuta Semen and Curcumae Tuber had a small or no effect on the viability of HepG2 cells. Conclusions: In the sovereign, minister, assistant and courier principle (君臣佐使論), Ginseng Radix (Red Ginseng) corresponds to the sovereign component because it provides cell protection effects, Angelica Gigantis Radix corresponds to minister medicinal because it kills cells, Schisandrae Fructus corresponds to the assistant medicinal to help red ginseng having cell protect effects. Cuscuta Semen and Curcumae Tuber correspond to the courier medicinal having no effect in cell viability in HepG2. We hope this study provides motivation for advanced research on the sovereign, minister, assistant and courier principle.

Antiproliferative and Antioxidative Activities of Methanol Extracts of Echinacea angustifolia (Echinacea angustifolia 메탄올 추출물의 암세포 증식억제 및 항산화 효과)

  • Lee Joon-Kyoung;Koo Seung-Ja
    • Korean journal of food and cookery science
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    • v.21 no.3 s.87
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    • pp.311-318
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    • 2005
  • Echinacea, also blown as the purple coneflower, is a herbal medicine that has been used for centuries, customarily as a treatment for the common cold, coughs, bronchitis, upper respiratory infections, and some inflammatory conditions. We investigated the effects of methanol extracts of Echinacea angustifolia on the cytotoxicity against cancer cells $(HepG_2,\;3LL,\;HL60,\;L1210)$ and antioxidative activity. From the test results, each part of Echinaceashowed a cytotoxic effect against the cancer cell lines, and this cytotoxic effect increased with increasing sample concentration. At 1.0 mg/mL concentration the relative cytotoxic activities of the flower bud, leaf, stern and root parts were $90.5\%,\;52.7\%,\;37.1\%\;and\;19.2\%$, respectively, in $HepG_2$ cells, and $75.5\%,\;93.3\%,\;81.2\%,\;and\;75.1\%$ respectively, in HL60 cells, as evaluated by MTT assay. $IC_{50}(50\%\;inhibitory\;concentration)$ of the methanol extracts of the Echinacea flower bud was 0.214 mg/mL on /$HepG_2$ cells, and that of the Echinacea leaf and root was 0.166 mg/mL and 0.210 mg/mL, respectively, on HL60 cells. After /$HepG_2$ cells were incubated for 6 days at $37^{\circ}C$ with various concentrations of each part, the cell number increased while the inhibition rate on the /$HepG_2$ cell growth decreased. The antioxidative activities of the flower bud, leaf, stem and root parts were $59.0\%$ (0.75 mg/mL), $80.76\%$ (0.5 mg/mL), $95.5\%$ (0.25mg/mL) and $98.15\%$ (0.25 mg/mL), respectively, as evaluated by electron donating ability. These results indicated that Echinacea angustifolia has strong anticancer and antioxidative effects in vitro.