Treatment with 5-azacytidine Increases UCP2 levels in Hep3B and HT-29 Cells but Not in HepG2 Cells

Abstract Background: The function of the uncoupling protein 2 (UCP2) is different for each cancer cell, and the mechanism of production is unclear. DNA methylation affects protein expression and is one factor that transforms normal cells into cancer cells. The hepatocellular carcinoma Hep3B and HepG2 cells and colorectal cancer HT-29 cells were treated with 5-azacytidine (5-aza), a DNA demethylation agent, to observe the modification of UCP2 production and the methylation degree in the UCP2 promoter region.Methods: Promoter basal activity and degree of UCP2 production were measured in Hep3B, HepG2, and HT-29 cells. In addition, methylation-specific PCR (MSP) was performed to investigate the degree of methylation in the UCP2 promoter region. The methylation region in the UCP2 promoter was confirmed based on bisulfite sequencing.Results: In Hep3B cells in which UCP2 mRNA was not transcribed, the promoter basal activity was significantly higher than in HT-29 or HepG2 cells in which UCP2 mRNA was transcribed. Treatment with 5-aza increased UCP2 expression in Hep3B and HT-29 cells; however, the expression in HepG2 cells was unchanged. The UCP2 promoter in Hep3B cells has numerous methylated regions compared with HT-29 and HepG2 cells.Conclusion: The results of the present study revealed that inhibition of UCP2 production in Hep3B cells was due to multiple methylation of the promoter region. Investigating the mechanism that induces UCP2 production in cancer cells is important to understand the function of UCP2, which could aid in cancer treatment.

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RAB9A Plays an Oncogenic Role in Human Liver Cancer Cells

BioMed Research International ◽

10.1155/2020/5691671 ◽

2020 ◽

Vol 2020 ◽

pp. 1-8

Author(s):

Pengfei Sun ◽

Lei Li ◽

Zhongchao Li

Keyword(s):

Liver Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Hepg2 Cells ◽

Human Liver ◽

Apoptotic Pathway ◽

Liver Cancer Cells ◽

Human Liver Cancer ◽

Hep3b Cells ◽

Human Liver Cancer Cells

Background. RAB9, as a member of the Rab GTPase family, is required for the transport of the mannose-6-phosphate receptor (MPR) from late endosomes to trans-Golgi network (TGN). However, the role of RAB9A in tumors, including liver cancer, is still unknown. Methods. We used pcDNA3.1 plasmid to upregulate the expression of RAB9A in Hep3b cells and used specific shRNA to downregulate the expression of RAB9A in HepG2 cells. Biological functions of RAB9A were performed by CCK-8 assay, colony formation assay, apoptosis analysis, transwell assays, and wound healing assays. Finally, an in-depth mechanism study was performed by western blot. Results. RAB9A promoted the proliferation and clonality of Hep3b and HepG2 cells. RAB9A also inhibited apoptosis and the activation of mitochondrial apoptotic pathway. In addition, RAB9A promoted the invasion and migration of Hep3b and HepG2 cells. Importantly, RAB9A activated the AKT/mTOR signaling pathway in human liver cancer cells. A double-effect inhibitor (BEZ235) significantly hindered the effect of RAB9A overexpression on the proliferation and invasion of Hep3b cells. Conclusion. Our data suggest that RAB9A plays a carcinogenic role in human liver cancer progression partially through AKT signaling pathways, suggesting that RAB9A may serve as a potential therapeutic target for liver cancer therapy.

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Growth-inhibitory Effect of the Solar Salt-Doenjang on Cancer Cells, AGS and HT-29

Journal of the Korean Society of Food Science and Nutrition ◽

10.3746/jkfn.2009.38.12.1664 ◽

2009 ◽

Vol 38 (12) ◽

pp. 1664-1671 ◽

Cited By ~ 10

Author(s):

Sun Mi Lee ◽

Hae Choon Chang

Keyword(s):

Cancer Cells ◽

Inhibitory Effect ◽

Growth Inhibitory Effect ◽

Solar Salt ◽

Growth Inhibitory ◽

Ht 29

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Inhibitory Effects of Flavonoids on Growth of HT-29 Human Colon Cancer Cells

Journal of the Korean Society of Food Science and Nutrition ◽

10.3746/jkfn.2015.44.3.338 ◽

2015 ◽

Vol 44 (3) ◽

pp. 338-346

Author(s):

Young Cho ◽

Mi-Yong Choi

Keyword(s):

Colon Cancer ◽

Cancer Cells ◽

Human Colon ◽

Colon Cancer Cells ◽

Inhibitory Effects ◽

Human Colon Cancer ◽

Human Colon Cancer Cells ◽

Ht 29

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SUV39H1-Mediated DNMT1 is Participated in the Epigenetic Regulation of Smad3 in Cervical Cancer

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200721110016 ◽

2020 ◽

Vol 20 ◽

Author(s):

Li Zhang ◽

Sijuan Tian ◽

Minyi Zhao ◽

Ting Yang ◽

Shimin Quan ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Lines ◽

Epigenetic Regulation ◽

Promoter Region ◽

Methylation Status ◽

Regulation Mechanism ◽

Methylation Specific Pcr ◽

Specific Pcr ◽

Immune Factor ◽

Cancer Tissues

Background: Smad3 is a pivotal intracellular mediator for participating in the activation of multiple immune signal pathway. Objective: The epigenetic regulation mechanism of the positive immune factor Smad3 in cervical cancer remains unknown. Therefore, the epigenetic regulation on Smad3 is investigated in this study. Methods: The methylation status of SMAD3 was detected by Methylation-specific PCR (MS-PCR) and Quantitative Methylation-specific PCR (MS-qPCR) in cervical cancer tissues and cell lines. The underlying molecular mechanisms of SUV39H1-DNMT1-Smad3 regulation was elucidated using cervical cancer cell lines containing siRNA or/and overexpression system. Confirmation of the regulation of DNMT1 by SUV39H1 used Chromatin immunoprecipitation-qPCR (ChIP-qPCR). The statistical methods used for comparing samples between groups were paired t tests and one-way ANOVAs. Results: H3K9me3 protein which regulated by SUV39H1 directly interacts with the DNMT1 promoter region to regulate its expression in cervical cancer cells, resulting in the reduce expression of the downstream target gene DNMT1. In addition, DNMT1 mediates the epigenetic modulation of the SMAD3 gene by directly binding to its promoter region. The depletion of DNMT1 effectively restores the expression of Smad3 in vitro. Moreover, in an in vivo assay, the expression profile of SUV39H1-DNMT1 was found to correlate with Smad3 expression in accordance with the expression at the cellular level. Notably, the promoter region of SMAD3 was hypermethylated in cervical cancer tissues, and this hypermethylation inhibits the subsequent gene expression. Conclusion: These results indicate that SUV39H1-DNMT1 is a crucial Smad3 regulatory axis in cervical cancer. SUV39H1-DNMT1 axis may provide a potential therapeutic target for the treatment of cervical cancer.

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Anticancer Activity of Platinum (II) Complex with 2-Benzoylpyridine by Induction of DNA Damage, S-Phase Arrest, and Apoptosis

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520619666191112114340 ◽

2020 ◽

Vol 20 (4) ◽

pp. 504-517

Author(s):

Yu-Lan Li ◽

Xin-Li Gan ◽

Rong-Ping Zhu ◽

Xuehong Wang ◽

Duan-Fang Liao ◽

...

Keyword(s):

Dna Damage ◽

B Cell ◽

Anticancer Activity ◽

Cancer Cells ◽

Hepg2 Cells ◽

Caspase 3 ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

S Phase ◽

Caspase 9

Objective: To overcome the disadvantages of cisplatin, numerous platinum (Pt) complexes have been prepared. However, the anticancer activity and mechanism of Pt(II) complexed with 2-benzoylpyridine [Pt(II)- Bpy]: [PtCl2(DMSO)L] (DMSO = dimethyl sulfoxide, L = 2-benzoylpyridine) in cancer cells remain unknown. Methods: Pt(II)-Bpy was synthesized and characterized by spectrum analysis. Its anticancer activity and underlying mechanisms were demonstrated at the cellular, molecular, and in vivo levels. Results: Pt(II)-Bpy inhibited tumor cell growth, especially HepG2 human liver cancer cells, with a halfmaximal inhibitory concentration of 9.8±0.5μM, but with low toxicity in HL-7702 normal liver cells. Pt(II)- Bpy induced DNA damage, which was demonstrated through a marked increase in the expression of cleavedpoly (ADP ribose) polymerase (PARP) and gamma-H2A histone family member X and a decrease in PARP expression. The interaction of Pt(II)-Bpy with DNA at the molecular level was most likely through an intercalation mechanism, which might be evidence of DNA damage. Pt(II)-Bpy initiated cell cycle arrest at the S phase in HepG2 cells. It also caused severe loss of the mitochondrial membrane potential; a decrease in the expression of caspase-9 and caspase-3; an increase in reactive oxygen species levels; the release of cytochrome c and apoptotic protease activation factor; and the activation of caspase-9 and caspase-3 in HepG2 cells, which in turn resulted in apoptosis. Meanwhile, changes in p53 and related proteins were observed including the upregulation of p53, the phosphorylation of p53, p21, B-cell lymphoma-2-associated X protein, and NOXA; and the downregulation of B-cell lymphoma 2. Moreover, Pt(II)-Bpy displayed marked inhibitory effects on tumor growth in the HepG2 nude mouse model. Conclusion: Pt(II)-Bpy is a potential candidate for cancer chemotherapy.

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Brassinin Inhibits Proliferation in Human Liver Cancer Cells via Mitochondrial Dysfunction

Cells ◽

10.3390/cells10020332 ◽

2021 ◽

Vol 10 (2) ◽

pp. 332

Author(s):

Taeyeon Hong ◽

Jiyeon Ham ◽

Jisoo Song ◽

Gwonhwa Song ◽

Whasun Lim

Keyword(s):

Mitochondrial Dysfunction ◽

Cancer Cells ◽

Cell Lines ◽

Mapk Pathway ◽

Mapk Signaling ◽

Nuclear Antigen ◽

Cruciferous Vegetable ◽

Antiproliferative Effects ◽

Hep3b Cells ◽

Human Liver Cancer Cells

Brassinin is a phytochemical derived from Chinese cabbage, a cruciferous vegetable. Brassinin has shown anticancer effects on prostate and colon cancer cells, among others. However, its mechanisms and effects on hepatocellular carcinoma (HCC) have not been elucidated yet. Our results confirmed that brassinin exerted antiproliferative effects by reducing proliferating cell nuclear antigen (PCNA) activity, a proliferation indicator and inducing cell cycle arrest in human HCC (Huh7 and Hep3B) cells. Brassinin also increased mitochondrial Ca2+ levels and depolarized the mitochondrial membrane in both Huh7 and Hep3B cells. Moreover, brassinin generated high amounts of reactive oxygen species (ROS) in both cell lines. The ROS scavenger N-acetyl-L-cysteine (NAC) inhibited this brassinin-induced ROS production. Brassinin also regulated the AKT and mitogen-activated protein kinases (MAPK) signaling pathways in Huh7 and Hep3B cells. Furthermore, co-administering brassinin and pharmacological inhibitors for JNK, ERK1/2 and P38 decreased cell proliferation in both HCC cell lines more than the pharmacological inhibitors alone. Collectively, our results demonstrated that brassinin exerts antiproliferative effects via mitochondrial dysfunction and MAPK pathway regulation on HCC cells.

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Echinacoside exerts anti-tumor activity via the miR-503-3p/TGF-β1/Smad aixs in liver cancer

Cancer Cell International ◽

10.1186/s12935-021-01890-3 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Wen Li ◽

Jing Zhou ◽

Yajie Zhang ◽

Jing Zhang ◽

Xue Li ◽

...

Keyword(s):

Liver Cancer ◽

Cancer Cells ◽

Hepg2 Cells ◽

Luciferase Reporter ◽

Therapeutic Effects ◽

Dependent Manner ◽

Expression Levels ◽

Liver Cancer Cells ◽

Anti Tumor Activity ◽

Tgf Β1

Abstract Background Echinacoside (ECH) is the main active ingredient of Cistanches Herba, which is known to have therapeutic effects on metastatic tumors. However, the effects of ECH on liver cancer are still unclear. This study was to investigate the effects of ECH on the aggression of liver cancer cells. Methods Two types of liver cancer cells Huh7 and HepG2 were treated with different doses of ECH at different times and gradients. MTT and colony formation assays were used to determine the effects of ECH on the viability of Huh7 and HepG2 cells. Transwell assays and flow cytometry assays were used to detect the effects of ECH treatment on the invasion, migration, apoptosis and cell cycle of Huh7 and HepG2 cells. Western blot analysis was used to detect the effects of ECH on the expression levels of TGF-β1, smad3, smad7, apoptosis-related proteins (Caspase-3, Caspase-8), and Cyto C in liver cancer cells. The relationship between miR-503-3p and TGF-β1 was detected using bioinformatics analysis and Luciferase reporter assay. Results The results showed that ECH inhibited the proliferation, invasion and migration of Huh7 and HepG2 cells in a dose- and time-dependent manner. Moreover, we found that ECH caused Huh7 and HepG2 cell apoptosis by blocking cells in S phase. Furthermore, the expression of miR-503-3p was found to be reduced in liver tumor tissues, but ECH treatment increased the expression of miR-503-3p in Huh7 and HepG2 cells. In addition, we found that TGF-β1 was identified as a potential target of miR-503-3p. ECH promoted the activation of the TGF-β1/Smad signaling pathway and increased the expression levels of Bax/Bcl-2. Moreover, ECH could trigger the release of mitochondrial Cyto C, and cause the reaction Caspases grade. Conclusions This study demonstrates that ECH exerts anti-tumor activity via the miR-503-3p/TGF-β1/Smad aixs in liver cancer, and provides a safe and effective anti-tumor agent for liver cancer.

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Folic Acid-Targeted Paclitaxel-Polymer Conjugates Exert Selective Cytotoxicity and Modulate Invasiveness of Colon Cancer Cells

Pharmaceutics ◽

10.3390/pharmaceutics13070929 ◽

2021 ◽

Vol 13 (7) ◽

pp. 929

Author(s):

Antonella Grigoletto ◽

Gabriele Martinez ◽

Daniela Gabbia ◽

Tommaso Tedeschini ◽

Michela Scaffidi ◽

...

Keyword(s):

Folic Acid ◽

Cancer Cells ◽

Pharmacokinetic Profile ◽

Strong Impact ◽

Passive Targeting ◽

Drug Conjugates ◽

Tumor Delivery ◽

The Right ◽

Polymer Drug Conjugates ◽

Ht 29

Although selective tumor delivery of anticancer drugs has been sought by exploiting either passive targeting or by ligand-mediated targeting, a selective anticancer therapy remains an unmet medical need. Despite the advances which have been achieved by nanomedicines, nanosystems such as polymer-drug conjugates still miss the goal of clinical efficacy. In this study, we demonstrated that polymer-drug conjugates require a thoroughly chemical design and the right targeting agent/polymer ratio to be selective and effective towards cancer cells. In particular, two PEG conjugates carrying paclitaxel and targeted with different folic acid (FA)/PEG ratios (one or three) were investigated. The cytotoxicity study in positive (HT-29) and negative (HCT-15) FA receptor (FR)-cell lines demonstrated that the conjugates with one or three FAs were 4- or 28-fold more active in HT-29 cells, respectively. The higher activity of the 3-FA conjugate was confirmed by its strong impact on cell cycle arrest. Furthermore, FA targeting had a clear effect on migration and invasiveness of HT-29 cells, which were significantly reduced by both conjugates. Interestingly, the 3-FA conjugate showed also an improved pharmacokinetic profile in mice. The results of this study indicate that thorough investigations are needed to optimize and tune drug delivery and achieve the desired selectivity and activity towards cancer cells.

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Rhodium(i) N-heterocyclic carbene complexes: synthesis and cytotoxic properties

New Journal of Chemistry ◽

10.1039/d1nj00144b ◽

2021 ◽

Vol 45 (11) ◽

pp. 5176-5183

Author(s):

Ichraf Slimani ◽

Serap Şahin-Bölükbaşı ◽

Mustafa Ulu ◽

Enes Evren ◽

Nevin Gürbüz ◽

...

Keyword(s):

Cancer Cells ◽

Mtt Assay ◽

Incubation Time ◽

Human Cancer ◽

Cytotoxic Activities ◽

Carbene Complexes ◽

Human Cancer Cells ◽

Benzimidazolium Salts ◽

Ht 29

A series of benzimidazolium salts and their [RhCl(NHC)(COD)] complexes were synthesized. All compounds were screened for in vitro cytotoxic activities against a panel of human cancer cells (HT-29 colon, Ishikawa endometrial, U-87 glioblastoma) using the MTT assay for 48 h incubation time.

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