Ropivacaine promotes apoptosis of hepatocellular carcinoma cells through damaging mitochondria and activating caspase-3 activity

Our preliminary screening identified an extract from the rhizome of Dioscorea tokoro, which strongly suppressed the proliferation of HepG2 hepatocellular carcinoma cells and inhibited autophagy. This study aimed to isolate active compounds from the rhizome of D. tokoro that exert antiproliferative effects and inhibit autophagy. The bioassay-guided fractionation of the active fraction led to the isolation of two spirostan-type steroidal saponins, dioscin (1) and yamogenin 3-O-α-l-rhamnopyranosyl (1→4)-O-α-l-rhamnopyranosyl(1→2)-β-d-glucopyranoside (2), and the frostane-type steroidal saponin protodioscin (3) from the n-BuOH fraction. Furthermore, acid hydrolysis of 1 and 2 produced the aglycones diosgenin (4) and yamogenin (5), respectively. Compounds 1–5 suppressed proliferation of HepG2 cells. The analysis of structure-activity relationships indicated that the 25(R)-conformation, structures with a sugar moiety, and the spirostan-type aglycone moiety contributed to antiproliferative activity. Analysis of autophagy-related proteins demonstrated that 1–3 clearly increased the levels of both LC3-II and p62, implying that 1–3 deregulate the autophagic pathway by blocking autophagic flux, which results in p62 and LC3-II accumulation. In contrast, 1–3 did not significantly affect caspase-3 activation and PARP cleavage, suggesting that the antiproliferative activity of 1–3 occurred independently of caspase-3-mediated apoptosis. In summary, our study showed that 1–3, active compounds in the rhizome of D. tokoro, suppressed cell proliferation and autophagy, and might be potential agents for autophagy research and cancer chemoprevention.

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Wogonin and fisetin induction of apoptosis through activation of caspase 3 cascade and alternative expression of p21 protein in hepatocellular carcinoma cells SK-HEP-1

Archives of Toxicology ◽

10.1007/s00204-002-0346-6 ◽

2002 ◽

Vol 76 (5-6) ◽

pp. 351-359 ◽

Cited By ~ 115

Author(s):

Yen-Chou Chen ◽

Shing-Chuan Shen ◽

Woan-Rouh Lee ◽

Hui-Yi Lin ◽

Ching-Huai Ko ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Caspase 3 ◽

Carcinoma Cells ◽

Hepatocellular Carcinoma Cells ◽

P21 Protein ◽

Alternative Expression ◽

Induction Of Apoptosis

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Effect of the BBC3 Gene on the Proliferation and Apoptosis of Hepatocellular Carcinoma Cells Through p53-Regulated Signaling

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2386 ◽

2021 ◽

Vol 11 (1) ◽

pp. 135-141

Author(s):

Sheng Zheng ◽

Hua Yang ◽

Yefei Chang ◽

Dan Zhao ◽

Juan Yang

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Proliferation ◽

Hepg2 Cell ◽

Hepg2 Cells ◽

Caspase 3 ◽

Carcinoma Cells ◽

Caspase 9 ◽

Hepatocellular Carcinoma Cells ◽

P53 Signaling ◽

Normal Human Liver

We examined the effect of the BBC3 gene on hyperplasia and apoptosis in HepG2 cells and its underlying mechanism. Quantitative RT-PCR was used to determine the level of BBC3 expression in HL-7702 normal human liver cells and four different hepatocellular carcinoma cell lines (HepG2, HuH-7, HCCLM3 and MHCC97H). Transfection was performed with Lipofectamine 2000 reagent and the transfectants were divided into three groups: pcDNA-BBC3 group (transfected BBC3 over-expressing plasmid), pcDNA-NC group (transfected empty plasmid), and a Ctrl group (not transfected). Quantitative RT-PCR and western blot analysis were used to measure BBC3 expression. The CCK-8 assay was used to determine the effect of BBC3 on HepG2 cell proliferation. Flow cytometry was used for testing the effect of overexpressing BBC3 on apoptosis in HepG2 cells. The levels of cleaved-Caspase-3 (C-Caspase-3), cleaved-Caspase-9 (C-Caspase-9), and proteins associated with the p53 signaling pathway were assessed by western blot analysis. The level of BBC3 mRNA in HL-7702 normal human liver cells was significantly higher compared with that in human hepatocellular carcinoma cells including HepG2, HuH-7, HCCLM3 and MHCC97H (P < 0.05). The lowest level of BBC3 mRNA was observed in HepG2 cells. The level of BBC3 mRNA and protein in HepG2 cells were significantly higher compared with that of the pcDNA-NC group following transfection with a BBC3 overexpressing plasmid. HepG2 cell proliferation in the pcDNA-NC group was higher compared with that of the pcDNA-BBC3-transfected group (P < 0.05). The apoptotic rate and levels of cleaved-Caspase-3, cleaved-Caspase-9, p53, phospho-p53, and p21 protein in cells were higher compared with that of the pcDNA-NC group. No change was observed in the pcDNA-NC and Ctrl groups. The BBC3 gene was down-regulated in hepatocellular carcinoma cells. HepG2 cell proliferation can be inhibited and HepG2 cell apoptosis can be induced by the overexpression of BBC3 through activation of the p53 signaling pathway.

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Effects of NK-104 on apoptosis and caspase-3 activity in hepatocellular carcinoma cells

Journal of Chinese Integrative Medicine ◽

10.3736/jcim20070313 ◽

2007 ◽

Vol 5 (3) ◽

pp. 298-301

Author(s):

Jurong Wang

Keyword(s):

Hepatocellular Carcinoma ◽

Caspase 3 ◽

Carcinoma Cells ◽

Hepatocellular Carcinoma Cells

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Artemisiae Iwayomogii Herba Inhibits Growth, Motility, and the PI3K/AKT/mTOR Signaling Pathway in Hepatocellular Carcinoma Cells

Planta Medica ◽

10.1055/a-1167-4284 ◽

2020 ◽

Vol 86 (10) ◽

pp. 717-727

Author(s):

Juyoung Kim ◽

Kyung Hee Jung ◽

Jin Gyu Choi ◽

Myung Sook Oh ◽

Soon-Sun Hong

Keyword(s):

Hepatocellular Carcinoma ◽

Caspase 3 ◽

Carcinoma Cell ◽

Ex Vivo ◽

Ethanol Extract ◽

Carcinoma Cells ◽

Terminal Deoxynucleotidyl Transferase ◽

Mtor Signaling Pathway ◽

Hepatocellular Carcinoma Cells ◽

Hepatocellular Carcinoma Cell

Abstract Artemisia gmelinii (Artemisia iwayomogi) has been used in traditional medicine to cure various infectious diseases such as cholecystitis, hepatitis, and jaundice. In this study, the Artemisiae Iwayomogii Herba ethanol extract was investigated for the ability to inhibit growth of hepatocellular carcinoma and its underlying mechanism involved. The antiproliferative effect of Artemisiae Iwayomogii Herba ethanol extract was evaluated using cell viability and proliferation assays. The effect of Artemisiae Iwayomogii Herba ethanol extract on apoptosis was measured using western blotting, terminal deoxynucleotidyl transferase-mediated dUTP-biotin end labeling staining, JC-1 staining, cytochrome c release, immunohistochemistry, and immunofluorescence in ex vivo mouse xenografts. Artemisiae Iwayomogii Herba ethanol extract inhibited hepatocellular carcinoma cell growth and proliferation in a dose-dependent manner. The apoptotic effect of Artemisiae Iwayomogii Herba ethanol extract was observed via increased levels of cleaved caspase-3 and cleaved PARP, as well as elevated numbers of terminal deoxynucleotidyl transferase-mediated dUTP-biotin end labeling-positive apoptotic cells. Artemisiae Iwayomogii Herba ethanol extract also decreased XIAP and Mcl-1 expression via loss of mitochondrial membrane potential. Additionally, Artemisiae Iwayomogii Herba ethanol extract inhibited hepatocellular carcinoma cell invasion and migration. In the ex vivo model, Artemisiae Iwayomogii Herba ethanol extract significantly inhibited tumor cell proliferation and increased the number of apoptotic cells with more activated cleaved caspase-3. A mechanistic study revealed that Artemisiae Iwayomogii Herba ethanol extract effectively suppressed the PI3K/AKT/mTOR signaling pathway in hepatocellular carcinoma cells. Our findings demonstrate that Artemisiae Iwayomogii Herba ethanol extract can efficiently induce apoptosis and inhibit the growth, migration, and invasion of human hepatocellular carcinoma cells, and simultaneously block PI3K/AKT/mTOR pathway. We therefore suggest Artemisiae Iwayomogii Herba ethanol extract as a novel natural agent for prevention and therapy of hepatocellular carcinoma.

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Dual induction of caspase 3- and transglutaminase-dependent apoptosis by acyclic retinoid in hepatocellular carcinoma cells

Molecular Cancer ◽

10.1186/1476-4598-10-4 ◽

2011 ◽

Vol 10 (1) ◽

pp. 4 ◽

Cited By ~ 22

Author(s):

Hideki Tatsukawa ◽

Tetsuro Sano ◽

Yayoi Fukaya ◽

Naoto Ishibashi ◽

Makiko Watanabe ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Caspase 3 ◽

Carcinoma Cells ◽

Hepatocellular Carcinoma Cells ◽

Acyclic Retinoid

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Targeting survivin with prodigiosin isolated from cell wall of Serratia marcescens induces apoptosis in hepatocellular carcinoma cells

Human & Experimental Toxicology ◽

10.1177/0960327116651122 ◽

2016 ◽

Vol 36 (4) ◽

pp. 402-411 ◽

Cited By ~ 15

Author(s):

RA Yenkejeh ◽

MR Sam ◽

M Esmaeillou

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Wall ◽

Growth Inhibition ◽

Hepg2 Cells ◽

Caspase 3 ◽

Survivin Expression ◽

Cell Number ◽

Carcinoma Cells ◽

Dependent Manner ◽

Hepatocellular Carcinoma Cells

Background: Abnormal activation of the Wnt/β-catenin signaling pathway increases survivin expression that is involved in hepatocarcinogenesis. Therefore, downregulation of survivin may provide an attractive strategy for treatment of hepatocellular carcinoma. In this regard, little is known about the anticancer effects of prodigiosin isolated from cell wall of Serratia marcescens on the survivin expression and induction of apoptosis in hepatocellular carcinoma cells. Methods: Human hepatocellular carcinoma (HepG2) cells were treated with 100-, 200-, 400-, and 600-nM prodigiosin after which morphology of cells, cell number, growth inhibition, survivin expression, caspase-3 activation, and apoptotic rate were evaluated by inverted microscope, hemocytometer, MTT assay, RT-PCR, fluorometric immunosorbent enzyme assay, and flow cytometric analysis, respectively. Results: Prodigiosin changed morphology of cells to apoptotic forms and disrupted cell connections. This compound significantly increased growth inhibition rate and decreased metabolic activity of HepG2 cells in a dose- and time-dependent manner. After 24-, 48-, and 72-h treatments with prodigiosin at concentrations ranging from 100 nM to 600 nM, growth inhibition rates were measured to be 1.5–10%, 24–47.5%, and 55.5–72.5%, respectively, compared to untreated cells. At the same conditions, metabolic activities were measured to be 91–83%, 74–53%, and 47–31% for indicated concentrations of prodigiosin, respectively, compared to untreated cells. We also found that treatment of HepG2 cells for 48 h decreased significantly cell number and survivin expression and increased caspase-3 activation in a dose-dependent manner. Specifically, treatment with 600-nM prodigiosin resulted in 77% decrease in cell number, 88.5% decrease in survivin messenger RNA level, and 330% increase in caspase-3 activation level compared to untreated cells. An increase in the number of apoptotic cells (late apoptosis) ranging from 36.9% to 97.4% was observed with increasing prodigiosin concentrations. Conclusion: From our data, prodigiosin is an attractive compound that turns the profile of high-level survivin expression in hepatocellular carcinoma cells into that of normal cells and may provide a novel approach to the hepatocellular carcinoma-targeted therapy.

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Curcumin inhibits proliferation of hepatocellular carcinoma cells through down regulation of DJ-1

Cancer Biomarkers ◽

10.3233/cbm-190427 ◽

2020 ◽

Vol 29 (1) ◽

pp. 1-8

Author(s):

Lina Han ◽

Yu Wang ◽

Shulun Sun

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Proliferation ◽

Cell Apoptosis ◽

Hepg2 Cells ◽

Caspase 3 ◽

Akt Signaling ◽

Carcinoma Cells ◽

Pten Expression ◽

Hepatocellular Carcinoma Cells ◽

Proliferation And Apoptosis

PTEN exerts tumor suppressor role through inhibiting PI3K/AKT signaling. DJ-1 plays an oncogenic role through negatively regulation of PTEN expression. Curcumin (Cur) is a phenolic compound extracted from a variety of plant roots, with multiple anti-tumor pharmacological effects. This study aims to investigate whether Cur plays a role in the regulation of DJ-1-PENT/PI3K/AKT signaling as well as the proliferation and apoptosis of hepatocellular carcinoma cells. Normal human hepatocyte HL-7702 and hepatocellular carcinoma cell lines SMMC-7721 and HepG2 were cultured followed by analysis of the expression of DJ-1 and PTEN. SMMC-7721 and HepG2 cells were treated with different concentrations of Cur (0, 5, 10 μM) followed by measuring cell proliferation by CCK-8, caspase-3 activity as well as DJ-1 expression by western blot. In addition, SMMC-7721 or HepG2 cells were divided into two groups: Cur+pcDNA3.1-Blank and Cur+pcDNA3.1-DJ-1 for analysis of the expression of DJ-1, PTEN and p-AKT, cell apoptosis and proliferation. Compared with HL-7702, SMMC-7721 and HepG2 cells displayed significantly higher DJ-1 expression and lower PTEN expression. Cur treatment significantly inhibited proliferation of SMMC-7721 and HepG2 cells, increased caspase-3 activity and downregulated DJ-1 expression. Transfection of pcDNA3.1-DJ-1 significantly increased DJ-1 and p-AKT expression, promoted cell proliferation, but decreased PTEN expression and cell apoptosis. In conclusion, Cur inhibits proliferation of hepatocellular carcinoma cells and PTEN/PI3K/AKT signaling pathway via the reduction of DJ-1 expression, which provides new insights to the anticancer effects of curcumin in hepatocellular carcinoma.

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