• Asian Pacific Journal of Cancer Prevention
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• v.17 no.11
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• pp.5031-5035
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• 2016

Docetaxel, recognized as a stabilizing microtubule agent, is frequently administrated as a first line treatment for prostate cancers. Due to high side effects of monotherapy, however, combinations with novel adjuvants have emerged as an alternative strategy in cancer therapy protocols. Here, we investigated the combined effects of stattic and docetaxel on the DU145 prostate cancer cell line. Cytotoxicity was evaluated by MTT assay. To understand molecular mechanisms of stattic action, apoptotic related genes including Bcl-2, Mcl-1, Survivin and Bax were evaluated by real-time RT-PCR. Alteration in the expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 genes and Bax/Bcl-2 ratio were investigated via the $2^{{\Delta}{\Delta}CT}$ method. The $IC_{50}$ values for docetaxel and stattic were $3.7{\pm}0.9nM$ and $4.6{\pm}0.8{\mu}M$, respectively. Evaluation of key gene expression levels revealed a noticeable decrease in antiapoptotic Bcl-2 and Mcl-1 along with an increase in pro-apoptotic Bax mRNA levels (p<0.05). Our results suggest that combination of a STAT3 inhibitor with doctaxel can be considered as a potent strategy for induction of apoptosis via increasing Bax mRNA expression.

• Journal of Environmental Health Sciences
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• v.29 no.2
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• pp.16-22
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• 2003

BBP (Butyl benzyl phthalate), a widely used plasticizer. can enter the food and environment as consequence of its manufacture, use, and disposal. BBP was found to be developmental and teratogenic or endocrine disrupting chemical in rats. The effects of BBP were investigated in female rats (P) and second generation (F1) via lactations. Sprague-Dawley were given BBP by oral administration at 0, 5, 10, 100, 1000 mg/kg on day 0 to 21 of lactation period. The results were as follows : At maternal findings, there were some significant changes (p<0.05) in relative organ weight, especially liver and uterus weight by BBP administration. In estrous cycle, high treated group was inclined to be proestrus or estrus compared to control group. BBP indues estrous cycle earlier than the control group. At fetal findings, there were some significant changes in relative liver and spleen weight, especially 100, 1000 mg/kg administered groups. The relative weight of ventral prostate was decreased, so it was represent to dose-response tendency. Parent rats (P) were detected monobenzyl phthalate (MBeP) 3.21~5.81 $\mu\textrm{g}$/ml in 100, 1000 mg/kg dose groups. MBeP of male and female fetuses (F1) were detected at the level of 1.21~2.63 $\mu\textrm{g}$/ml of serum. Male serum concentration oi MBeP was higher than the females'. Estrogen receptor $\alpha$ expression by BBP and bisphenol A in uterus and testis of F1 were studied. The ER$\alpha$ expression were increased in F1 male testis and female uterus. F1 male showed distint ER$\alpha$ expression, especially in the combined exposrue. Synergistic ER$\alpha$ expression was found by combined treatment group of BBP and bisphenol A. From the above results, it could be concluded that the effects of dams and F1 by BBP administration during lactation period were estrogenic, and BBP can transfer to F1 via lactation, and make estrogenic at F1 reproductive organs.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6869-6874
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• 2013

Background: Vascular endothelial growth factor and matrix metalloproteinases are two important factors for angiogenesis associated with breast cancer growth and progression. The present study was aimed to examine the effects of tamoxifen and tranilast drugs singly or in combination on proliferation of breast cancer cells and also to evaluate VEGF and MMP-9 expression and VEGF secretion levels. Materials and Methods: Human breast cancer cell lines, MCF-7 and MDA-MB-231, were treated with tamoxifen and/or tranilast alone or in combination and percentage cell survival and proliferative activity were evaluated using LDH leakage and MTT assays. mRNA expression and protein levels were examined by real-time RT-PCR and ELISA assay, respectively. Results: LDH and MTT assays showed that the combined treatment of tamoxifen and tranilast resulted in a significant decrease in cell viability and cell proliferation compared with tamoxifen or tranilast treatment alone, with significant decrease in VEGF mRNA and protein levels. We also found that tamoxifen as a single agent rarely increased MMP-9 expression. A decrease in MMP-9 expression was seen after treatment with tranilast alone and in the combined treatment MMP-9 mRNA level was decreased. Conclusions: This combination treatment can able to inhibit growth, proliferation and angiogenesis of breast cancer cells.

• Korean Journal of Ecology and Environment
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• v.38 no.4 s.114
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• pp.510-517
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• 2005

The filamentous cyanobacterium Anabaena is capable of both photosynthesis and nitrogen fixation which probably facilitated its incredible adaptation and proliferation in freshwater environments. A small gene, patS, was found to block nitrogen fixing cells from developing which resulted in death of Anabaena in the absence of combined nitrogen sources. We analyzed the DNA sequences in the vicinity of the patS gene by using a codon usage program and detected no codon bias other than the patS open reading frame. Three overlapping cosmids that contain the patS gene were identified, and the presence of other known heterocyst-controlling genes was examined. The patS expression in response to nitrogen starvation was analyzed at the level of transcription and translation by using Northern blot analyses and lacZ-reporter-gene fusion experiments, respectively. The patS expression increased rapidly (within 12 hours) upon the removal of combined nitrogen from the media.

• Korean Journal of Veterinary Research
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• v.49 no.1
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• pp.1-8
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• 2009

The ubiquitin-proteasome mediated protein degradation pathway plays an important role in regulating both cell proliferation and cell death. Proteasome inhibitors are well known to induce apoptosis in various human cancer cell lines. We investigated the effect of combined treatment with proteasome inhibitor and TRAIL, and a possible mechanism of the enhancing apoptosis by the both treatment, on TRAIL-resistant non-small cell lung cancer. A549 cells were exposed to the N-Acetyl-Leu-Leu-Norleu-al (ALLN) as a proteasome inhibitor and then treated with recombinant TRAIL protein. In A549 cells under proteasome inhibition conditions by pretreatment with ALLN, TRAIL treatment significantly decreased cell viability compared to that ALLN and TRAIL alone treatment. Also, the both treatment induced cell damage through DNA fragmentation and p53 expression. In addition, the combined treatment of both markedly increased caspase-8 activation, especially the exposure for 2 h, and Bax expression and induced the dissipation of mitochondrial transmembrane potential in A549 cells. Taken together, these findings showed that proteasome inhibition by ALLN enhanced TRAIL-induced apoptosis via DNA degradation by activated P53 and mitochondrial transmembrane potential loss by caspase-8 activation and bax expression. Therefore, our results suggest that proteasome inhibitor may be used a very effectively chemotherapeutic agent for the tumor treatment, especially TRAIL-resistant tumor cell.

• Journal of Korean Dental Science
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• v.7 no.2
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• pp.58-65
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• 2014

Purpose: This study was performed to evaluate the effect of chitosan combined with absorbable gelatin compressed sponge on the expression of osteoblastic differentiation marker genes during the healing of rat extraction socket. Materials and Methods: Twenty-four male Wistar rats were used. In control group, the extraction socket was closed with suture. In chitosan group, the socket was filled with chitosan combined with Gelfoam (Pharmacia & Upjohn Co.) and closed with suture. In each group, the animals were sacrificed at 3 days, 1 week, 2 weeks, and 4 weeks postoperatively. The expression of osteoblastic differentiation marker genes, including BSP, OCN, Runx2, and Col1 were quantified by real-time polymerase chain reaction. Result: Compared to control group, the mRNA level of BSP in chitosan group increased significantly at 2 weeks after extraction and the level of OCN decreased significantly at 3 days and 4 weeks after extraction (P<0.05). The mRNA levels of OCN, Runx2, and Col1 in chitosan group increased slightly at 2 weeks after extraction, but there was no statistical difference between groups. Conclusion: The results indicate that chitosan has some effects on the expression of osteogenic genes during the healing of extraction sockets.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.6
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• pp.468-473
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• 2005

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy worldwide. The molecular mechanisms involved in the development and progression of these carcinomas are not well known. Abnormalities of genomic methylation patterns have been attributed a role in carcinogenesis and local de novo methylation at tumor suppressor loci was held to be involved in silencing of tumor suppressor genes. Using Ms APPCR, we previously isolated a hypermethylated fragment corresponded to the 5' end of TPEF gene from primary liver and lung cancer cells. To confirm the inactivation of TPEF gene by hypermethylation in HNSCC, we investigated correlation between methylation pattern and expression of TPEF in 10 HNSCC cell lines. In methylation analysis such as combined-bisulfite restriction analysis(COBRA) and bisulfite sequencing, only RPMI 2650 showed none methylated pattern and another 9 cell lines showed dense methylation. The TPEF gene expression level analysis using RT-PCR showed that these 9 cell lines had not or significantly low expression levels of TPEF as compared with RPMI 2650. In addition, the increase of TPEF reexpression by 5-AzaC as demethylating agent in 9 cell lines also indicated that TPEF expression was regulated by hypermethylation. These results of this study demonstrate that epigenetic silencing of TPEF gene by aberrant methylation could play an important role in HNSCC carcinogenesis.

• Journal of Bone Metabolism
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• v.25 no.4
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• pp.251-266
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• 2018

Background: The causal networks among genes that are commonly expressed in osteoblasts and during bone metastasis (BM) of breast cancer (BC) are not well understood. Here, we developed a machine learning method to obtain a plausible causal network of genes that are commonly expressed during BM and in osteoblasts in BC. Methods: We selected BC genes that are commonly expressed during BM and in osteoblasts from the Gene Expression Omnibus database. Bayesian Network Inference with Java Objects (Banjo) was used to obtain the Bayesian network. Genes registered as BC related genes were included as candidate genes in the implementation of Banjo. Next, we obtained the Bayesian structure and assessed the prediction rate for BM, conditional independence among nodes, and causality among nodes. Furthermore, we reported the maximum relative risks (RRs) of combined gene expression of the genes in the model. Results: We mechanistically identified 33 significantly related and plausibly involved genes in the development of BC BM. Further model evaluations showed that 16 genes were enough for a model to be statistically significant in terms of maximum likelihood of the causal Bayesian networks (CBNs) and for correct prediction of BM of BC. Maximum RRs of combined gene expression patterns showed that the expression levels of UBIAD1, HEBP1, BTNL8, TSPO, PSAT1, and ZFP36L2 significantly affected development of BM from BC. Conclusions: The CBN structure can be used as a reasonable inference network for accurately predicting BM in BC.

• Anatomy and Cell Biology
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• v.57 no.1
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• pp.85-96
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• 2024

Glioblastoma is the most common primary malignant brain tumor in adults. Temozolomide (TMZ) is an FDA-approved drug used to treat this type of cancer. Cinnamaldehyde (CIN) is a derivative of cinnamon extract and makes up 99% of it. The aim of this study was to investigate the in vitro combined effect of CIN and TMZ on human glioblastoma multiforme T98G cell line viability. In this study, we used 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-tertazolium bromide (MTT) method to evaluate the extent of IC₅₀, acridine orange, Giemsa and Hoechst staining to evaluate the manner of apoptosis and the Western blotting method to examine the expression change of apoptotic proteins. Our results show that TMZ has an inhibitory effect on CIN when both used in combination at concentrations of 300 and 100 µM (P<0.05) and has a cytotoxic effect when used alone at the same concentrations (P<0.05). The western blotting result showed that TMZ at concentrations of 2,000 and 1,000 µM significantly increased Bax expression and decreased Bcl2 expression (P<0.05), indicating that TMZ induced apoptosis through the mitochondrial pathway. However, CIN had no effect on Bax and Bcl2 expressions, thus causing apoptosis from another pathway. Also, the Bax:Bcl2 expression ratio at concentrations combined was lower than that for TMZ 1,000 µM and higher than that for CIN 150 and 100 µM (P<0.05), which confirms the inhibitory effect of TMZ on CIN. From the present study, we conclude that TMZ in combination with CIN has an inhibitory effect on increasing the cytotoxicity rate.

• Nutritional Sciences
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• v.8 no.3
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• pp.159-168
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• 2005

Phytoestrogens, especially Yak-kong or soybean-derived isoflavones have been traditionally used as a supplement of estrogen for preventing postmemopausal osteoporosis in oriental folk medicine. In our previous study, the treatment of Yak-kong and soybean increased estrogen receptor-a (ERa) expression and proliferation of MG-63 osteoblastic cells. In contrast, the increase of estrogen receptor-$\beta$ (ER$\beta$) expression in proliferating MG-63 cells with Yak-kong and soybean treatment was less pronounced, which suggested that ER$\beta$ may play a role rather in the regulation of bone cell differentiation To determine the role of ER$\beta$ in Yak-kong or soybean mediated regulation of bone cell differentiation, we established MG-63 cell lines stably expressing either ER$\beta$ or antisense ER$\beta$ RNAs. Increased expression of ER$\beta$ did not affect ERa expression and proliferation of MG-63 cells. However, increased expression of ER$\beta$ in MG-63 cells (ER$\beta$-MG63 cells) selectively enhanced Yak-kong or soybean induced expression of osteoprotegerin (OPG), a novel soluble glycoprotein which is secreted from osteoblasts and mediates the signal for osteoclast differentiation. Inhibition of ER$\beta$ expression by antisense ER$\beta$ RNAs (As-ER$\beta$-MG63) caused these cells to insensitize Yak-kong or soybean induced expression of OPG but increased MG-63 cell proliferation. Furthermore, the comparable effects between Yak-kong and the combined treatment of genistein and daidzein at $0.5{\times}l0^{-8}$ M, which is a concentration of these two isoflavones similar to Yak-kong at 0.001 mg/mL, on OPG expression in ER$\beta$-MG63 cell demonstrate that the enhanced expression of OPG with Yak-kong treatment is mediated by the synergistic effect of low leveled isoflavones in the extracts. Together, coupled with low level of ER expression in osteoclasts, our data demonstrate that ER$\beta$ in osteoblasts plays an important role in Yak-kong and soybean mediated inhibition of osteoclast differentiation indirectly by enhancing the expression of OPG.