2.Experimental study of inhibition of tumor cell proliferation by a novel gene SPATA12.
Zhiwen LIU ; Yiting LIN ; Xuanming LIU ; Weiwei YU ; Yunsheng ZHANG ; Dan LI
Journal of Central South University(Medical Sciences) 2012;37(3):222-227
OBJECTIVE:
To explore the inhibitory role of spermatogenesis-associated gene 12 (SPATA12) on tumor cell proliferation and its possible mechanism.
METHODS:
The expression pattern of SPATA12 in testicular tumors was investigated by in situ hybridization analysis using tissue microarrays. The effects of SPATA12 on tumor cell proliferation and colony formation was detected by 3-(4.5-dimethylthiazol-2- yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and colonyforming assays, respectively. The changes of expression level of cell cycle genes in tumor cells were detected by reverse transcription polymerase chain reaction(RTPCR).
RESULTS:
In situ hybridization analysis showed that the SPATA12 was highly expressed in normal adult testis, but lacking in testicular tumors such as seminoma. MTT assay and colony-forming assay indicated that the exogenous expression of SPATA12 could suppress both tumor cell proliferation and colony formation. RT-PCR showed that the expression of cyclin A1 gene was markedly suppressed and the level of cyclin D1 was somewhat reduced following SPATA12 transfection. However, no obvious changes were observed in mRNA expression of cyclin B1 or cyclin E1 after SPATA12 transfection.
CONCLUSION
SPATA12 could be an inhibitor during the development of tumor via regulation of cell cycle genes.
Cell Line, Tumor
;
Cell Proliferation
;
Genes, Tumor Suppressor
;
Genes, cdc
;
HeLa Cells
;
Homeodomain Proteins
;
genetics
;
metabolism
;
Humans
;
Male
;
Testicular Neoplasms
;
genetics
;
pathology
;
Transfection
3.Preferential Cytotoxic Effect of Genistein on G361 Melanoma Cells Via Inhibition of the Expression of Focal Adhesion Kinase.
Sang Rye PARK ; Hyun Ho KWAK ; Bong Soo PARK ; Gyoo Cheon KIM
International Journal of Oral Biology 2012;37(4):189-195
Resistance to the induction of apoptosis is a possible mechanism by which tumor cells can survive anti-neoplastic treatments. Melanoma is notoriously resistant to anti-neoplastic therapy. Previous studies have demonstrated focal adhesion kinase (FAK) overexpression in melanoma cell lines. Given its probable role in mediating resistance to apoptosis, many researchers have sought to determine whether the downregulation of FAK in melanoma cells would confer a greater sensitivity to anti-neoplastic agents. Genistein is a known inhibitor of protein-tyrosine kinase (PTK), which may attenuate the growth of cancer cells by inhibiting the PTK-mediated signaling pathway. This present study was undertaken to investigate the effect of genistein on the expression of FAK and cell cycle related proteins in the G361 melanoma cell line. Genistein was found to have a preferential cytotoxic effect on G361 melanoma cells over HaCaT normal keratinocytes. Genistein decreased the expression of 125 kDa phosphotyrosine kinase and the FAK protein in particular. Genistein treatment did not affect the expression of p53 in G361 cells in which p21 is upregulated. The expression of cyclin B and cdc2 was downregulated by genistein treatment. Taken together, our data indicate that genistein induces the decreased proliferation of G361 melanoma cells via the inhibition of FAK expression and regulation of cell cycle genes. This suggests that the use of genistein may be a viable approach to future melanoma treatments.
Apoptosis
;
Cell Cycle
;
Cell Line
;
Cyclin B
;
Down-Regulation
;
Focal Adhesion Protein-Tyrosine Kinases
;
Focal Adhesions
;
Genes, cdc
;
Genistein
;
Keratinocytes
;
Melanoma
;
Negotiating
;
Phosphotransferases
;
Phosphotyrosine
;
Protein-Tyrosine Kinases
;
Proteins
4.RNA-sequencing Profiles of Cell Cycle–Related Genes Upregulated during the G2-Phase in Giardia lamblia
Juri KIM ; Mee Young SHIN ; Soon Jung PARK
The Korean Journal of Parasitology 2019;57(2):185-189
To identify the component(s) involved in cell cycle control in the protozoan Giardia lamblia, cells arrested at the G1/S- or G2-phase by treatment with nocodazole and aphidicolin were prepared from the synchronized cell cultures. RNA-sequencing analysis of the 2 stages of Giardia cell cycle identified several cell cycle genes that were up-regulated at the G2-phase. Transcriptome analysis of cells in 2 distinct cell cycle stages of G. lamblia confirmed previously reported components of cell cycle (PcnA, cyclin B, and CDK) and identified additional cell cycle components (NEKs, Mad2, spindle pole protein, and CDC14A). This result indicates that the cell cycle machinery operates in this protozoan, one of the earliest diverging eukaryotic lineages.
Aphidicolin
;
Cell Culture Techniques
;
Cell Cycle
;
Cell Cycle Checkpoints
;
Cyclin B
;
Gene Expression Profiling
;
Genes, cdc
;
Giardia lamblia
;
Giardia
;
Nocodazole
;
Spindle Poles
5.The role of BRCA1 in DNA damage response.
Jiaxue WU ; Lin-Yu LU ; Xiaochun YU
Protein & Cell 2010;1(2):117-123
BRCA1 is a well-established tumor suppressor gene, which is frequently mutated in familial breast and ovarian cancers. The gene product of BRCA1 functions in a number of cellular pathways that maintain genomic stability, including DNA damage-induced cell cycle checkpoint activation, DNA damage repair, protein ubiquitination, chromatin remodeling, as well as transcriptional regulation and apoptosis. In this review, we discuss recent advances regarding our understanding of the role of BRCA1 in tumor suppression and DNA damage response, including DNA damage-induced cell cycle checkpoint activation and DNA damage repair.
Apoptosis
;
genetics
;
BRCA1 Protein
;
genetics
;
physiology
;
Breast Neoplasms
;
genetics
;
DNA Damage
;
genetics
;
DNA Repair
;
genetics
;
Female
;
Genes, Tumor Suppressor
;
Genes, cdc
;
physiology
;
Humans
;
Mutation
;
Ovarian Neoplasms
;
genetics
;
Signal Transduction
;
genetics
6.Effects of Postaglandin E1 on the Oxidative Stress Injury Index and the Expression of PCNA in Vein Grafts of Rabbit Carotid Arteries.
Tae Seung LEE ; In Mok JUNG ; Jung Yun CHOI ; Myung Hee CHUNG ; Jeong Wook SEO ; Jongwon HA ; Sang Joon KIM
Journal of the Korean Surgical Society 2000;58(2):161-170
BACKGROUND: This study investigated the protective mechanism of Prostagladin E1 (PGE1) against intimal hyperplasia after vein interposition grafts in rabbits. It has been demonstrated that active oxygen species contribute to vascular smooth muscle cell growth via early cell cycle gene activation. We attempted to study whether PGE1 had an effect on the inhibition of the oxidative stress injury index (8-OHdG, MDA). METHODS: Forty-eight jugular vein grafts were inserted into the carotid arteries of male hyperlipidemic New Zealand white rabbits, which were divided into 2 groups (saline group and PGE1 group). Saline and Prostaglandin E1 (0.1 microgram/kg/min) were administered as a continuous infusion for 2 hours every day from just before graft interposition to harvest. The vein grafts were harvested at 6 hour, 1 day, 1 week, and 2 week after grafting and rapidly stored in liquid nitrogen ( 70oC). 8-OHdG was measured by using high performance liquid chromatography coupled with electrochemical detection (HPLC-EC), and malondialdehyde (MDA) was measured by using thiobarbituric acid (TBA) assay. PC 10 index and intimal thickness of the grafts were measured with a computer digitalized image analyzer. RESULTS: There was no difference in 8-OHdG levels between the saline and the PGE1 groups. PGE1 had more inhibitory effect on the MDA level as an oxidative stress injury index, but its action was restricted to 1 day. A morphometric analysis and an immunohistochemical study showed that the PGE1 group had more suppressive effects both in intimal thickeness and proliferating cell nuclear antigen (PCNA) expression than the saline group (p<0.05). CONCLUSION: These results suggest that PGE1 is effective in preventing intimal hyperplasia after vein interposition grafts in rabbits and may play a role in inhibiting oxidative stress injury.
Alprostadil
;
Carotid Arteries*
;
Chromatography, Liquid
;
Genes, cdc
;
Humans
;
Hyperplasia
;
Jugular Veins
;
Male
;
Malondialdehyde
;
Muscle, Smooth, Vascular
;
Nitrogen
;
Oxidative Stress*
;
Proliferating Cell Nuclear Antigen*
;
Rabbits
;
Reactive Oxygen Species
;
Transplants*
;
Veins*
7.DNA repair and synthetic lethality.
Gong-She GUO ; Feng-Mei ZHANG ; Rui-Jie GAO ; Robert DELSITE ; Zhi-Hui FENG ; Simon N POWELL
International Journal of Oral Science 2011;3(4):176-179
Tumors often have DNA repair defects, suggesting additional inhibition of other DNA repair pathways in tumors may lead to synthetic lethality. Accumulating data demonstrate that DNA repair-defective tumors, in particular homologous recombination (HR), are highly sensitive to DNA-damaging agents. Thus, HR-defective tumors exhibit potential vulnerability to the synthetic lethality approach, which may lead to new therapeutic strategies. It is well known that poly (adenosine diphosphate (ADP)-ribose) polymerase (PARP) inhibitors show the synthetically lethal effect in tumors defective in BRCA1 or BRCA2 genes encoded proteins that are required for efficient HR. In this review, we summarize the strategies of targeting DNA repair pathways and other DNA metabolic functions to cause synthetic lethality in HR-defective tumor cells.
Animals
;
Antineoplastic Agents
;
pharmacology
;
Breast Neoplasms
;
genetics
;
DNA Repair
;
drug effects
;
genetics
;
Gene Expression Regulation, Neoplastic
;
drug effects
;
Genes, Lethal
;
genetics
;
Genes, Tumor Suppressor
;
drug effects
;
Genes, cdc
;
drug effects
;
Humans
;
Mutagenesis
;
Poly(ADP-ribose) Polymerase Inhibitors
;
Rad52 DNA Repair and Recombination Protein
;
antagonists & inhibitors
;
Recombination, Genetic
;
drug effects
;
genetics
8.The effects of nonyl phenoxypolyethoxyl ethanol on cell damage pathway gene expression in SK-NSH cells.
Samel PARK ; Il Woong HWANG ; Jin Sheon KIM ; Hyo Chul KANG ; Su Yeon PARK ; Hyo Wook GIL ; Ho Yeon SONG ; Sae Yong HONG
The Korean Journal of Internal Medicine 2015;30(6):873-883
BACKGROUND/AIMS: Most pesticide formulations contain both chief and additive ingredients. But, the additives may not have been tested as thoroughly as the chief ingredients. The surfactant, nonyl phenoxypolyethoxylethanol (NP40), is an additive frequently present in pesticide formulations. We investigated the effects of NP40 and other constituents of a validamycin pesticide formulation on cell viability and on the expression of genes involved in cell damage pathways. METHODS: The effects of validamycin pesticide ingredients on cell viability and of NP40 on the mRNA expression of 80 genes involved in nine key cellular pathways were examined in the human neuroblastoma SK-N-SH cell line. RESULTS: The chemicals present in the validamycin pesticide formulation were cytotoxic to SK-N-SH cells and NP40 showed the greatest cytotoxicity. A range of gene expression changes were identified, with both up- and down-regulation of genes within the same pathway. However, all genes tested in the necrosis signaling pathway were down-regulated and all genes tested in the cell cycle checkpoint/arrest pathway were up-regulated. The median fold-change in gene expression was significantly higher in the cell cycle checkpoint/arrest pathway than in the hypoxia pathway category (p = 0.0064). The 70 kDa heat shock protein 4 gene, within the heat shock protein/unfolded protein response category, showed the highest individual increase in expression (26.1-fold). CONCLUSIONS: NP40 appeared to be particularly harmful, inducing gene expression changes that indicated genotoxicity, activation of the cell death (necrosis signaling) pathway, and induction of the 70 kDa heat shock protein 4 gene.
Aged
;
Cell Cycle Checkpoints/drug effects/genetics
;
Cell Line, Tumor
;
Cell Survival/drug effects
;
Dose-Response Relationship, Drug
;
Female
;
Gene Expression Regulation/drug effects
;
Genes, cdc
;
HSP110 Heat-Shock Proteins/genetics/metabolism
;
Humans
;
Inositol/*analogs & derivatives/chemistry/poisoning
;
Necrosis
;
Neurons/*drug effects/metabolism/pathology
;
Nonoxynol/chemistry/*toxicity
;
Pesticides/chemistry/*poisoning
;
RNA, Messenger/metabolism
;
Signal Transduction/drug effects
;
Surface-Active Agents/chemistry/*toxicity
9.Investigation on the molecular mechanisms of anti-hepatocarcinoma herbs of traditional Chinese medicine by cell cycle microarray.
Guang-Liang WANG ; Cheng-Bin CHEN ; Jian-Ming GAO ; Hong NI ; Tong-Shun WANG ; Li CHEN
China Journal of Chinese Materia Medica 2005;30(1):50-54
OBJECTIVETo design DNA microarray and investigate the molecular anti-tumor mechanism of herbs of traditional Chinese medicine.
METHODcDNA microarrays consisting of 56 probes representing 24 human cell cycle genes were constructed, Four anti-hepatocarcinoma herbs including Radix Linderae, Hebra Artemisiae Annuae, Radix Amebiae, Radix Astragli, were chosen. Effects of herbs on SMMC-7721 cell cycle were observed by flow cytometry assay. Effects of herbs on cell cycle gene expression in SMMC-7721 cells were analyzed by comparing hybridization of Dig-Labeled cDNAs from herb-treated cells and cDNAs from untreated cells.
RESULTExpressions of cell cycle geneswere changed in different degrees after herbs treated. Some genes were down-regulated and some genes were up-regulated. The changes in gene expression agreed with the results of flow cytometry assay.
CONCLUSIONThe results suggest that these herbs may have effects on cell cycle and DNA damage checkpoint genes which may be the mechanism of the herbs, and DNA microarray can be used to investigate the biological function of extracts of traditional Chinese medicine.
Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Artemisia ; chemistry ; Astragalus membranaceus ; chemistry ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Cyclin-Dependent Kinases ; genetics ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gene Amplification ; Gene Expression Profiling ; Genes, cdc ; drug effects ; Humans ; Lindera ; chemistry ; Lithospermum ; chemistry ; Liver Neoplasms ; metabolism ; pathology ; Oligonucleotide Array Sequence Analysis ; methods ; Plants, Medicinal ; chemistry ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; cdc25 Phosphatases ; genetics ; metabolism