• 한국식품영양학회지
• /
• 제35권6호
• /
• pp.464-472
• /
• 2022

The present study was designed to investigate the antiproliferative activity and molecular mechanisms of Bibimbap in HT-29 human colorectal adenocarcinoma cells. Bibimbap extract inhibited the proliferation of HT-29 cells by 50% at a concentration of 10.1±0.17 mg/mL for 48 h. The population of live cells decreased slightly, and the morphology changed with a reduction in cell volume (pyknosis) with Bibimbap. Treatment with 5 mg/mL of Bibimbap resulted in slight cell shrinkage. Furthermore, as the Bibimbap dose increased to 10 mg/mL, these characteristics were more evident, and HT-29 cells exhibited partial detachment by staining with the DNA-binding dye Hoechst 33342. Flow cytometric analysis by Annexin V and PI double staining showed that Bibimbap increased the levels of apoptosis. Analysis of the mechanism of these events showed that Bibimbap-treated cells exhibited a mitochondria-dependent apoptotic pathway through the modulation of caspase-3, caspase-8, caspase-9, and poly-ADP ribose polymerase, as well as Bax and Bcl-2 expression in dose- and time-dependent manners. Consequently, Bibimbap exerts a significant antiproliferative effect on HT-29 human colorectal adenocarcinoma cells.

• 한국식품영양과학회지
• /
• 제44권4호
• /
• pp.475-482
• /
• 2015

본 연구에서는 STZ 유발 당뇨 쥐에서 고혈당 및 산화적 스트레스 관련 간 손상 기전에 oligonol의 영향에 대해 살펴보고자 하였다. 10일 동안 oligonol을 투여한 결과 20 mg/kg oligonol 투여군에서 당뇨대조군에 비해 간 무게 및 간조직의 glucose, ROS, peroxynitrite, 지질과산화물 농도가 유의적으로 감소하였다. 간조직의 항산화 효소 단백질 발현을 측정한 결과 정상군에 비해 당뇨대조군에서 발현 정도가 낮아졌지만 oligonol 투여군에서는 유의적으로 증가하였다. 또한 caspase-3 효소의 단백질 발현은 당뇨대조군에서 증가하였으나 oligonol 투여군에서 그 발현이 억제되었다. 따라서 oligonol의 투여는 STZ-유발 당뇨 모델에서 ROS 및 지질과산화물 생성 억제와 항산화효소 작용의 증가 및 세포 사멸 작용이 있는 caspase-3 발현 억제를 통해 고혈당 및 산화적 스트레스에 의해 유발되는 간 손상에 대한 보호 효과가 있는 것으로 사료된다.

• 동의생리병리학회지
• /
• 제21권1호
• /
• pp.93-97
• /
• 2007

We previously reported the anti-proliferative effect of Chungjogupae-tang (CJGPT) in human lung carcinoma A549 cells, which was associated with the induction of cyclin-dependent kinase inhibitor p21 in a tumor suppressor p53-independent manner. CJGPT treatment also resulted in the inhibition of prostaglandin E2 release A549 cells by the down-regulation of cyclooxygenase-2. In the present study, we investigated the pathway of the induction of apoptotic cell death by CJGPT in A549 cells. It was found that CJGPT could inhibit the cell viability and induce the apoptotic cell death of A549 cells in a dose-dependent manner as measured by hemocytometer counts, flow cytometry analysis and agarose gel electrophoresis. Apoptosis of A549 cells by CJGPT was associated with a down-regulation of anti-apoptotic Bcl-2 and inhibitor of apoptosis proteins (IAPs) expression. Additionally, DNA fragmentation by CJGPT was connected with the activation of inhibitor of caspase-activated DNase/DNA fragmentation factor 45 (ICAD/DFF45) protein expression.

• 동의생리병리학회지
• /
• 제26권2호
• /
• pp.199-205
• /
• 2012

An extract of Alnus japonica (Betulaceae) cortex has been traditionally used for purifying blood, and curing feces containing blood, enteritis, diarrhea, alcoholism and cut wounds. In the present study, we demonstrated that the ethanol extract of Alnus japonica (EAJ) exhibited significantly cytotoxicity in human colon adenocarcinoma HT-29 cells. The results showed that the induction of apoptosis in HT-29 cells by EAJ was characterized by chromatin condensation and activation of caspase-3. EAJ-induced activation of caspase-9 and -3 caused the cleavage of poly ADP-ribose polymerase (PARP) and the release of cytochrome c. The expressions of Bcl-2 and Bid were reduced by EAJ in HT-29 cells, whereas pro-apoptotic protein Bak was increased in the cells. EAJ-induced, dose-dependent induction of apoptosis was accompanied by sustained phosphorylation of MAP kinases (JNK and p38 MAPK), ASK1, and p53. NAC administration, a scavenger of ROS, reversed EAJ-induced cell death. In conclusion, these results indicated that EAJ can cause apoptosis through a ROS-mitochondria-caspases-dependent pathway in human HT-29 cells.

• 대한한방부인과학회지
• /
• 제19권4호
• /
• pp.47-60
• /
• 2006

Purpose : This study was conducted to investigate the inhibitory effects of Scutellaria barbata D. D on on the cell proliferation of HeLa Cells. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Scutellaria barbata D. D on solution for 24, 48 and 72 hours for the direct inhibitory effects of Scutellaria barbata D. D on. Then we examined the effect of Scutellaria barbata D. D on solution on the cell proliferation inhibition by XTT assay. DNA fragmentation, MAP kinase activity and caspase activity by FACS analysis in HeLa cells. Results : We found that the proliferation of HeLa cells was significantly decreased in Scutellaria barbata D. D on solution containing groups comparing with a control group in a concentration-dependant manner. When HeLa cells were cultivated for 24 hours with 5% Scutellaria barbata D. D on solution containing group, the percentage of HeLa cells with activated caspase was the highest. Scutellaria barbata D. D on solution reduced the MAP kinase activity of HeLa cells comparing with the control group. By the XTT assay, the cell's activity was decreased in 5% and 10% Scutellaria barbata D. D on solution containing groups in 24 and 72 hours cultivation and 10% group in 48 hours. DNA fragmentation and caspase-3 activity of HeLa cells, however, were changed insignificantly. Conclusion : From this study we could suggest that Scutellaria barbata D. D on is available to the inhibition and apoptosis of human cervical carcinoma cell line, HeLa cells in vitro.

• 대한의생명과학회지
• /
• 제11권4호
• /
• pp.481-486
• /
• 2005

This study was performed to define roles of telomerase and apoptosis and their relationships with clinicopathologic characteristics in colorectal cancers. We performed TRAP (Telomeric Repeat Amplification Protoco1)-ELISA assay for telomerase activity and immunohistochemistry of active caspase-3 expression for apoptosis in 35 colorectal cancers. Increased telomerase activity was detected in $71.4\%$ (25/35) and average apoptotic index was 14.6 per 1000 tumor cells. Telomerase activity and caspase 3 expression had no significant association with clinicopathological characteristics, however, increased telomerase activity was more frequently found in progressed colorectal cancers. Although there is no definitive relation, low apoptotic index group was more frequent in cases with increased telomerase activity. These date indicate that telomerase might be involved in progression of colorectal cancers. We suggest that there is a need for further study to define the relationship between telomerase and apoptosis in colorectal cancers.

• 동의생리병리학회지
• /
• 제25권5호
• /
• pp.816-822
• /
• 2011

Kaempferol, a component of Polygonati rhizoma, is one of the herbal flavonoids, which is used in therapeutic agent for anti-hypercholesterol, anti-hypertension and anti-diabetes. And it is also known to be effective in anti-cancer therapy for breast, prostate and other type of cancers. However, the anti-cancer therapeutic mechanisms are pooly understood. To address molecular mechanism underlying kaempferol-induced anti-cancer effects, we determined the effect of kaempferol on cell growth of the lung cancer cell lines, A549, H1299 and H460. From the FACS analysis, measurement of caspase activity, DAPI and tryptophan blue staining, and DNA fragmentation assay, we found that kaempferol induces apoptosis and H460 cells are most sensitive among the tested cell lines. In addition, we performed microarray to identify the genome-wide expression profiling regulated by kaempferol. Lots of cell cycle-related genes were under-expressed, whereas the genes related to TGF-beta/SMAD pathway were over-expressed in kaempferol-treated H460 cells. Additionally, kaempferol also increased expression levels of apoptosis related genes such as death receptors, FAS, TRAIL-R and TNF-R, and casepase-8 and caspase-10. Overall, our results suggest that kaempferol promotes anti-lung cancer therapeutic effects by inducing G1 arrest and apoptosis through TGF-beta/SMAD pathway and death receptors/caspase pathway, respectively.

• Natural Product Sciences
• /
• 제26권3호
• /
• pp.191-199
• /
• 2020

Fucoidan, a natural component of brown seaweed, has various biological activities such as anti-cancer activity, anti-oxidant, and anti-inflammatory against various cancer cells. However, the fucoidan has been implicated in melanoma cells via apoptosis signaling pathway. Therefore, we investigated apoptosis with fucoidan in A2058 human melanoma cells with dose- and time-dependent manners. In our results, A2058 cells viability decreased at relatively short-time and low-concentration through fucoidan. This effects of fucoidan on A2058 cells appeared to be mediated by the induction of apoptosis, as manifested by morphological changes through DNA-binding dye Hoechst 33342 staining. When a dose of 80 ㎍/mL fucoidan was treated, the cells were observed: crescent or ring-like structure, chromatin condensation, and nuclear fragmentation. With the increase at 100 ㎍/mL fucoidan, the cell membrane is intact throughout the total process, including membrane blebbing and loss of membrane integrity as well as increase of sub-G1 DNA. Furthermore, to understand the exact mechanism of fucoidan-treated in A2058 cells, western blotting was performed to detect apoptosis-related protein expression. In this study, Bcl-2 family proteins can be regulated by fucoidan, suggesting that fucoidan-induced apoptosis is modulated by intrinsic pathway. Therefore, expression of Bcl-2 and Bax may result in altered permeability, activating caspase-3 and caspase-9. And the cleaved form of poly ADP-ribose polymerase was detected in fucoidan-treated A2058 cells. These results suggest that A2058 cells are highly sensitive to growth inhibition by fucoidan via apoptosis, as evidenced by activation of extracellular signal-regulated kinases/p38/Bcl-2 family signaling, as well as alteration in caspase-9 and caspase-3.

• Asian Pacific Journal of Cancer Prevention
• /
• 제17권7호
• /
• pp.3281-3287
• /
• 2016

Bioactive compounds extracted from leaves and twigs of Goniothalamus griffithii include pinocembrin (PCN) and goniothalamin (GTN). The objectives of this study were to investigate the cytotoxic activities of PCN and GTN and their influence on molecular signaling for cell death in several human cancer cell lines compared to normal murine fibroblast NIH3T3 cells. GTN exhibited the most potent cytotoxicity against MCF-7 > HeLa > HepG2 > NIH3T3 cells with $IC_{50}$ values of 7.33, 14.8, 37.1 and $65.4{\mu}M$, respectively, whereas PCN was cytotoxic only to HepG2 cells with $IC_{50}$ values of ${\sim}80{\mu}M$. Apoptotic cell death was confirmed by staining the cells with annexin V-FITC and propidium iodide (PI) employing flow cytometry. Apoptosis was shown by externalization of phosphatidylserine in goniothalamin-treated MCF-7 cells in a dose response manner. Positive PI-stained cells with the typical morphology of apoptotic cells were increased dose-dependently. Furthermore, reduction of mitochondrial transmembrane potential was found in goniothalamin-treated MCF-7, HepG2 and HeLa cells. GTN treatment in MCF-7 increased caspase-3, -8 and -9 activities while GTN-induced HeLa cells showed an increase of both caspase-3 and -9 activities. But an increased caspase-8 activity was demonstrated in GTN- and PCN-treated MCF-7 and HepG2 cells, respectively. Taken together, GTN- and PCN-induced human cancer cell apoptosis was through different molecular mechanisms or signaling pathways, which might be due to different machineries in different types of cancer cells, as evidenced by the compound-modulated caspase activities in both intrinsic and/or extrinsic pathways.

• 대한한방부인과학회지
• /
• 제19권2호
• /
• pp.77-91
• /
• 2006

Purpose : To address the ability of Dohaekseungkitang (DST: a commonly used herb formulation in Korea, Japan and China to have anti-cancer effect on cervical carcinoma), we investigated the effects of DST on programmed cell death (apoptosis) in HeLa human cervical carcinoma cells. Methods : We cultured HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% CO2. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : After the treatment of DST for 48 hours, apoptosis occurred in a dose-dependent manner. In this study, we have shown that DST induces calpain and the associated caspase-8 and -9 activations. Apoptosis was prevented by pre-incubation of the cells with the calcium cHeLator-BAPTA-AM, calcium channel blocker-Nif edipine or Ryonidine agonist-Ryonidine peptide, implicating calcium in the apoptotic process. Ubiquitous calpains (mu- and m-calpain) have been repeatedly implicated in apoptosis, especially in calcium-related apoptosis. However this study showed 1hat either calpain inhibitor-calpastin or caspase-3 inhibitor-DEVD- did not blocked the herb formulation-induced apoptosis in HeLa human cervical carcinoma cells. D ST initiates a cell death pathway that is partially dependent of caspases. DST-induced apoptosis requires caspase-independent mechanism. Conclusion : We conclude that DST-induced calpain activation triggers the intrinsic apoptotic pathway in which caspase-independent mechanism is also involved.