OBJECTIVETo investigate the effects of snakegourd root polysaccharide on apoptosis of human breast cancer cells (MCF-7 cells).

METHODSColorimetric MTT assay was used to measure the inhibition of snakegourd root polysaccharide on MCF-7 cells. The morphological changes of MCF-7 cells were observed by fluorescence microscope after DAPI staining and transmission electron microscope. The apoptosis of MCF-7 cells was examined by DNA agarose gel electrophoresis analysis of DNA fragmentation amd flow cytometry. The activity of Caspase-3 and Caspase-8 was detected by colorimetric assay.

RESULTSPolysaccharide of snakegourd root significantly inhibited MCF-7 cells in a dose-and time-dependent manner. The nuclear condensation and marginalization were observed by DAPI staining and transmission electron microscope. The characteristic ladder of apoptosis in DNA electrophoresis was detected in MCF-7 cells treated with 10.0 μmol/L polysaccharide of snakegourd root at d 2. The activities of Caspase-3 and Caspase-8 were increased in a time-dependent manner. The rates of apoptosis in MCF-7 cells were (5.2 ±1.3)%, (13.1 ±4.7)%, (27.6 ±6.8)% and (43.8 ±9.8)% treated with 1.0,5.0,10.0 and 20.0 μmol/L snakegourd root polysaccharide at d 2,respectively. The maximal activities of intracellular Caspase-3 and Caspase-8 were (2.32 ±0.12)U/μg and (1.92 ±0.11)U/μg at d 2 and d 1, respectively when MCF-7 cells were treated with 10.0 μmol/L.

CONCLUSIONThe polysaccharide of snakegourd root can induce the apoptosis of MCF-7 cells,which is associated with the activation of intracellular Caspase-3 and Caspase-8.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Humans ; MCF-7 Cells ; Plant Roots ; chemistry ; Polysaccharides ; pharmacology ; Trichosanthes ; chemistry

PURPOSE : Several lines of evidence have indicated that deregulation of apoptosis is involved in the mechanism of cancer development. Caspase-8 activation plays a central role in the initiation phase of apoptosis, while caspase-7 is one of the main execution phase caspases of apoptosis. The aim of this study was to explore the possibility that genetic alterations of the caspase-8 and caspase-7 genes are involved in the development of human non-small cell lung cancer (NSCLC). MATERIALS AND METHODS : We have analyzed the entire coding region of both the caspase-7 and caspase-8 genes to detect the somatic mutations in 100 NSCLCs by using polymerase chain reaction (PCR)- single strand conformation polymorphism (SSCP). RESULTS : The PCR-SSCP analysis detected no mutations in the entire coding regions of both the caspase-7 and caspase-8 genes in the NSCLCs. CONCLUSION : The data presented here suggests that both the caspase-7 and caspase-8 genes may not be somatically mutated in human NSCLCs
Apoptosis ; Carcinoma, Non-Small-Cell Lung* ; Caspase 7* ; Caspase 8 ; Caspases ; Clinical Coding ; Humans ; Polymerase Chain Reaction

Today, many materials as drug are developed having various prominent function in order to treatment of disease or cancer. Among these materials, especially docosahexaenoic acid (DHA), main constituents of omega-3 fatty acid, has a lot of beneficial and natural effects, so it has been known as anticancer material especially breast cancer. Breast cancer is disease taking high occurrence level among feminine diseases. DHA has anticancer effects on breast cancer cell, representatively inducing apoptosis, inhibiting proliferation or metastasis. Main effect of DHA on breast cancer cell is apoptosis inducing, which has mechanism that treated DHA causes lipid peroxidation increasing reactive oxygen species (ROS) level and it activates caspase 8 and caspase 9 so activated caspase occurs apoptosis. Cell lines of breast cancer are MDA-MB-231, MCF-7, SK-BR-3, T47D and ZR75. Especially this article uses the MCF-7 cell line at experiment of anti-proliferation by DHA, the MDA-MB-231 cell line at experiment of anti-metastasis by DHA, because that cell line has specialized metastasis activity. Therefore, this paper discusses the effects of natural material DHA as drug of breast cancer.
Apoptosis ; Breast Neoplasms* ; Breast* ; Caspase 8 ; Caspase 9 ; Cell Line ; Lipid Peroxidation ; MCF-7 Cells ; Neoplasm Metastasis ; Reactive Oxygen Species

6-Gingerol exerts anti-tumor effects in various cancer cell models. We evaluated the effect of 6-gingerol on the growth of MCF-7 breast cancer cells and MCF-10A breast epithelial cells to determine whether any growth-inhibitory effects found were attributable to apoptosis, and to elucidate the underlying mechanism of action. 6-Gingerol inhibited the viability of both cell lines in a dose- and time-dependent manner; however, the degree of inhibition was greater in MCF-7 than MCF-10A cells. By flow cytometry, induction of dose- and time-dependent apoptosis was found, and the magnitude of apoptosis was also markedly greater in MCF-7 than MCF-10A cells. Expression of caspase-3 and poly (ADP-ribose) polymerase (PARP) was observed in MCF-7 cells treated with 6-gingerol, and further cleavage of PARP occurred in these cells. We suggest that 6-gingerol induces apoptosis in human breast cancer cells mainly by promoting caspase-3 expression and subsequent degradation of PARP.
Apoptosis ; Breast Neoplasms* ; Breast* ; Caspase 3 ; Cell Line ; Epithelial Cells ; Flow Cytometry ; Humans* ; MCF-7 Cells

Although much information has been accumulated about the synergistic interaction of proteasome inhibitors and HDAC inhibitors to induce apoptosis in a certain type of cells, much less is known currently about the underlying mechanism. This study was undertaken to explore the combination effect of a histone deacetylase inhibitor, TSA, and a proteasome inhibitor, lactacystin, on the induction of apoptosis. Pretreatment of TSA and subsequent treatment of lactacystin showed the strong antitumor activity and nuclear condensation. Western blot assay showed that combination treatment of TSA and lactacystin increased Bax/Bcl-2 ratio and decreased level of XIAP. Activation of caspase-7 and cleavage of PARP were demonstrated after the combination treatment. In combination treatment group, cell cycle arrest was induced at G2/M phase and abolished increase in proteasome activity. This study is elucidating the mechanims whereby targeting apoptotic machineries may help in directing therapeutic strategies.
Apoptosis ; Blotting, Western ; Caspase 7 ; Cell Cycle Checkpoints ; Histone Deacetylase Inhibitors* ; Histone Deacetylases* ; Histones* ; MCF-7 Cells* ; Proteasome Endopeptidase Complex* ; Proteasome Inhibitors*

An FDA approved drug for the treatment of type II diabetes, Troglitazone (TRO), a peroxisome proliferator–activated receptor gamma agonist, is withdrawn due to severe idiosyncratic hepatotoxicity. In the search for new applications of TRO, we investigated the cellular effects of TRO on YD15 tongue carcinoma cells. TRO suppressed the growth of YD15 cells in the MTT assay. The inhibition of cell growth was accompanied by the induction of cell cycle arrest at G₀/G₁ and apoptosis, which are confirmed by flow cytometry and western blotting. TRO also suppressed the expression of cell cycle proteins such as cyclin D1, cdk2, cdk4, cyclin B1, cdk1(or cdc2), cyclin E1 and cyclin A. The inhibition of cell cycle proteins was coincident with the up-regulation of p21(CIP1/WAF1) and p27(KIP1). In addition, TRO induces the activation of caspase-3 and caspase-7, as well as the cleavage of PARP. Further, TRO suppressed the expressions of Bcl-2 without affecting the expressions of Bad and Bax. Overall, our data supports that TRO induces cell cycle arrest and apoptosis on YD15 cells.
Apoptosis ; Blotting, Western ; Caspase 3 ; Caspase 7 ; Cell Cycle Checkpoints ; Cell Cycle Proteins ; Cyclin A ; Cyclin B1 ; Cyclin D1 ; Cyclins ; Flow Cytometry ; Peroxisomes ; Tongue* ; Up-Regulation

Plant-derived triterpenoids commonly possesses biological properties such as anti-inflammatory, anti-microbial, anti-viral and anti-cancer. Luvunga scandens is one of the plant that produced triterpenoids. The aims of the study was to analyze cell cycle profile and to determine the expression of p53 unregulated modulator of apoptosis (PUMA), caspase-8 and caspase-9 genes at mRNA level in MCF-7 cell line treated with two triterpenoids, flindissol (1) and 3-oxotirucalla-7,24-dien-21-oic-acid (2) isolated from L. scandens. The compounds were tested for cell cycle analysis using flow cytometer and mRNA expression level using quantitative RT-PCR. The number of MCF-7 cells population which distributed in Sub G1 phase after treated with compound 1 and 2 were 7.7 and 9.3% respectively. The evaluation of the expression of genes showed that both compounds exhibited high level of expression of PUMA, caspase-8 and caspase-9 as normalized to β-actin via activation of those genes. In summary, the isolated compounds of L. scandens plant showed promising anticancer properties in MCF-7 cell lines.
Apoptosis ; Caspase 8 ; Caspase 9 ; Cell Cycle Checkpoints* ; Cell Cycle* ; Flow Cytometry ; G1 Phase ; Gene Expression* ; MCF-7 Cells* ; Plants ; Puma ; RNA, Messenger

OBJECTIVETo design hammerhead ribozymes against mouse caspase-7 and to study their expression and cleavage activity in vitro.

METHODSThe secondary structures of ribozyme and caspase-7 genes were analyzed and simulated by computer. Ribozymes DNA sequences were synthesized by automatic synthetic apparatus. Caspase-7 DNA sequence was acquired by reverse transcription PCR. Ribozymes and caspase-7 DNA sequences were separately cloned into pBSKneo U6 and pGEM-T vectors. Ribozymes and caspase-7 mRNA were obtained by transcription in vitro, and ribozymes cleavage activity was identified by cleavage experiment in vitro.

RESULTSTwo ribozymes named Rz333 and Rz394 targeting 333 and 394 sites in caspase-7 mRNA were designed by computer software, and their DNA sequences were synthesized. The expression vector of caspase-7 and plasmids containing Rz333 and Rz394 were reconstructed successfully. Ribozymes and caspase-7 mRNA were expressed by in vitro transcription. In vitro cleavage experiments showed that Rz333 cleaved caspase-7 mRNA and produced 243nt and 744nt segments. The cleavage efficiency is 67.98%, while Rz394 cannot cleave caspase-7 mRNA.

CONCLUSIONSRz333 can site-specifically cleave caspase-7 mRNA.

Animals ; Base Sequence ; Caspase 7 ; Caspase Inhibitors ; Caspases ; genetics ; Cloning, Molecular ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; RNA, Catalytic ; biosynthesis ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics

To investigate apoptosis of mouse leukemia cell (WEHI-3) induced by econazole and its mechanism, apoptosis induced by econazole was examined by flow cytometry, while free calcium ([Ca(2+)]i) was determined by Fura-2 fluorescein load technique. The protein was isolated from endoplasmic reticulum of WEHI-3 cells, and then the expression of caspase-12 and caspase-7 was evaluated by Western blot. The results showed that WEHI-3 exhibited typical change of apoptosis when it was treated by econazole, [Ca(2+)]i was significantly higher in comparison with the control. The expression of caspase-12 and caspase-7 enhanced as the econazole concentration increased. In conclusion, econazole can induce WEHI-3 cell apoptosis and the caspase-12 plays a key role in this process.
Animals ; Apoptosis ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Caspase 12 ; metabolism ; Caspase 7 ; metabolism ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Econazole ; pharmacology ; Flow Cytometry ; Leukemia ; metabolism ; pathology

Recent years, the incidence and mortality of prostate cancer have increased dramatically in China. At earlier stages, most diagnosed prostate cancers are responsive to androgen depletion treatment, yet, nearly all patients will eventually progress to metastatic androgen-independent prostate cancer (AIPC), which still has no effective therapeutic method or drug to deal with. 11'-Deoxyverticillin A (C42) belongs to the family of epipolythiodioxopiperazines (ETPs), an interesting class of fungal toxins that inhibit farnesyl transferase. Compounds holding such a property have been explored as putative anticancer agents. In this study, using PC3M cells, an AIPC cell line, we investigated the effect of the compound on apoptosis and explored the underlying mechanism. It revealed that C42 markedly enhanced the activity of caspase-3/7 and increased the accumulation of the cleaved PARP, all of which are the markers of apoptosis. It also revealed that C42 either decreased cell viability or inhibited the growth of PC3M cells. Moreover, we observed that the loss of cell viability and cell growth inhibition induced by C42 were both time- and dosage dependent. Taken together, we indicated that C42 can induce caspase-dependent apoptosis in AIPC cells, and the results presented here will broaden our knowledge about the molecular mechanisms by which C42 exerts its anticancer activity, and future work in this direction may provide valuable information in the development of these compounds into effective cancer therapeutic strategies against androgen-independent prostate cancer.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 7 ; metabolism ; Cell Line, Tumor ; Disulfides ; pharmacology ; Farnesyltranstransferase ; antagonists & inhibitors ; Humans ; Male ; Mycotoxins ; pharmacology ; Piperazines ; pharmacology ; Prostatic Neoplasms ; pathology