• Biomolecules & Therapeutics
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• 제32권1호
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• pp.104-114
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• 2024

Licochalcone C (LCC; PubChem CID:9840805), a chalcone compound originating from the root of Glycyrrhiza inflata, has shown anticancer activity against skin cancer, esophageal squamous cell carcinoma, and oral squamous cell carcinoma. However, the therapeutic potential of LCC in treating colorectal cancer (CRC) and its underlying molecular mechanisms remain unclear. Chemotherapy for CRC is challenging because of the development of drug resistance. In this study, we examined the antiproliferative activity of LCC in human colorectal carcinoma HCT116 cells, oxaliplatin (Ox) sensitive and Ox-resistant HCT116 cells (HCT116-OxR). LCC significantly and selectively inhibited the growth of HCT116 and HCT116-OxR cells. An in vitro kinase assay showed that LCC inhibited the kinase activities of EGFR and AKT. Molecular docking simulations using AutoDock Vina indicated that LCC could be in ATP-binding pockets. Decreased phosphorylation of EGFR and AKT was observed in the LCC-treated cells. In addition, LCC induced cell cycle arrest by modulating the expression of cell cycle regulators p21, p27, cyclin B1, and cdc2. LCC treatment induced ROS generation in CRC cells, and the ROS induction was accompanied by the phosphorylation of JNK and p38 kinases. Moreover, LCC dysregulated mitochondrial membrane potential (MMP), and the disruption of MMP resulted in the release of cytochrome c into the cytoplasm and activation of caspases to execute apoptosis. Overall, LCC showed anticancer activity against both Ox-sensitive and Ox-resistant CRC cells by targeting EGFR and AKT, inducing ROS generation and disrupting MMP. Thus, LCC may be potential therapeutic agents for the treatment of Ox-resistant CRC cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제27권4호
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• pp.314-320
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• 2001

Matrix metalloproteinases have long been viewed as ideal candidates for proteinases that enables tumor cells to permeated basement membrane defenses and invade surrounding tissue. There is growing evidence that the MMPs have an expanded role, as they are important for the creation and maintenance of a microenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. MT-MMPs are not secreted but instead remaining attached to cell surfaces. Although not all of the MT-MMPs are fully characterized, MT-MMPs have important role in localizing and activating secreted MMPs. The MMP genes are transcriptionally responsive to a wide variety of oncogene, growth factors, cytokine, and hormones. Currently, a number of MMP inhibitors are being developed and some have reached clinical trials as anti-metastatic or anti-cancer therapies. MT1-MMP is involved in the activation of proMMP-2. MT1-MMP is significant not only as a tumor marker but as a new target for chemotherapy against cancer. The purpose of this study was to evaluate the effects of protein kinase C inhibitor(genistein) on the proliferation of HT1080 and expression of MT1-MMP mRNA. Human fibrosarcoma cell line HT1080 was cultured and divided 2 groups. The experimental group was treated with $100{\mu}M$ genistein and incubated 12h, 24h for $[3^H]-thymidine$ uptake assay and northern hybridization individually. And the control group was treated with same amount of PBS for the above procedures. $[3^H]-thymidine$ incorporation was measured with ${\beta}$ ray detector. And RT-PCR and northern blotting for MT1-MMP mRNA was performed. The results were as follows 1. $[3^H]-thymidine$ uptake was reduced in experimental group with statistical significance. 2. MT1-MMP mRNA expression was significantly reduced in experimental group. These results showed that protein kinase C inhibitor (genistein) inhibited proliferation of HT1080 and almost completely blocked transcription of MT1-MMP mRNA. So, it is possible to use the protein kinase inhibitor (genistein) as anti-metastatic and anti-proliferative agent.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5535-5538
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• 2012

Background: There have been case reports of oral squamous cell carcinoma arising from gingival overgrowth induced by phenytoin - an antiepileptic drug. However, a detailed analysis for the presence of mutations in p53 and ras genes, which are the two most frequently mutated genes in cancers, in phenytoin induced gingival overgrowth tissues has hitherto not been performed. Methods: Cellular DNA isolated from twenty gingival overgrowth tissues collected from patients undergoing phenytoin therapy were amplified using primers for p53 (exons 5-8) and H-ras (exons 1-2) genes. The PCR amplicons were then gel purified and subjected to direct sequencing analysis to screen for mutations. Results: Direct sequencing of twenty samples of phenytoin induced gingival growth did not identify mutations in any of the exons of p53 and H-ras genes that were analyzed. Conclusion: Our result indicates that mutational alteration of p53 and H-ras genes is infrequent in phenytoin induced gingival growth, which thus suggests a non malignant nature of this pathology. The findings in the present study are clinically significant as a large number of epileptic patients are treated with phenytoin.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권3호
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• pp.218-228
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• 2010

발암물질로 알려진 Hydroquinone (HQ)은 치과용 합성수지를 구성하는 중요한 성분으로서 지금까지 치과용 재료영역에서 널리 사용되고 있으며 구강 내에서 HQ의 유출이 일어나는 것으로 확인 되었다. 따라서 구강암의 기원이 되는 인체상피세포의 발암화에 HQ가 미치는 영향을 평가하였다. HQ에 의한 인체세포 독성을 평가하기위해 LDH assay를 실시하고 세포 독성이 높지 않은 용량을 실험 용량으로 설정하였다. 인체 세포의 발암화를 평가하기 위해 세포 발암화 지표로서 cell saturation density, soft-agar colony formation 및 cell aggregation의 분석을 사용한 결과 고용량인 50 ${\mu}M$을 제외한 모든 용량에서 발암화 지표의 변화를 나타내지 않아 HQ의 발암력은 매우 낮은 것으로 추정된다. 그러나 발암촉진제인 TPA와 함께 투여 시 발암력의 증가를 보여 주변 환경의 여건에 따라 발암력이 증가할 수 있음을 입증하였다. HQ를 노출 후 세포사멸화를 측정하기 위해 DNA fragmentation변화를 분석한 결과 10 ${\mu}M$부터 50 ${\mu}M$까지 노출 시간 의존형의 증가를 나타내었으며 50 ${\mu}M$과 같은 고용량 농도에서는 노출시간 의존적 세포사멸 효과를 보였다. 따라서 세포 발암화를 일으킨 용량에서 세포사멸도 함께 일어나 HQ에 의한 발암화에 세포사멸이 관여함을 보였다. HQ는 ROS를 생성하였으며 Trolox, NAC와 같은 항산화물에 의한 ROS의 차단 효과와 BSO와 같은 GSH 고갈 유발 물질에 따른 ROS의 급격한 증가는 HQ가 인체세포에서 ROS를 효율적으로 생성함을 입증하는 결과이다. 세포간의 신호전달기작 조절에 중요한 역할을 하는 효소인 protein kinase C (PKC)를 immunoblot으로 분석한 결과 PKC-${\alpha}$의 활성이 증가 된 반면 PKC-${\beta}II$의 영향은 나타나지 않았다. 따라서 특정 이성질체에 대한 특이적인 효소반응이 발암화에 관여할 것으로 추정된다. 본 연구결과 치과용 합성수지 구성성분인 HQ 유출에 따른 인체상피세포의 발암성은 매우 낮은 것으로 추정되나 발암촉진제 등과의 상호작용에 의한 발암성 증가는 HQ의 구강암 발생 평가에 고려되어야 할 사항이다. 따라서 본 연구는 구강암의 예방을 위한 과학적인 접근 방법 및 기반 자료를 제시하였고 치과용 합성수지사용의 적정성에 대한 과학적인 판단을 할 수 있는 근거를 제공 하였다. 또한 본 연구 결과는 새로운 치과용 합성수지 개발의 필요성 및 개선방향을 제시 할 수 있는 근거로 활용될 수 있을 것으로 사료된다.

• Journal of Oral Medicine and Pain
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• 제38권3호
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• pp.221-233
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• 2013

담즙산은 지방의 흡수와 콜레스테롤의 항상성을 조절하는 유전자의 전사에 관여하는 필수 콜레스테롤의 생성물이다. 담즙산 합성유도체인 HS-1200이 여러 가지 암세포에서 세포자멸사(apoptosis)를 유도한다는 것이 알려져 있다. 본 연구는 사람혀 편평세포암종세포(SCC25 cells)에서 담즙산 합성유도체인 HS-1200과 대표적인 항암제인 cisplatin의 병용처리 후 세포자멸사 증가효과가 있는지 알아보기 위해 수행하였다. HS-1200과 cisplatin의 병용처리가 단독처리에 비해서 효과적인 세포생존율 감소가 있는지 확인하기 위해서 MTT법을 시행하였고, 세포자멸사의 유도와 증가를 알기 위해서는 DNA 전기영동법, Hoechst 염색법, DNA hypoploidy법 을 사용하였다. 그리고 세포자멸사에 관계하는 단백질의 발현 변화와 세포내에서의 이동을 밝혀내기 위해서 Western blot 분석과 면역형광 염색법을 수행하였다. 더 나아가서 proteasome 활성도와 사립체막 전위 변화를 측정하였다. 본 연구에서는 HS-1200과 cisplatin을 병용처리한 SCC25 세포에서 핵의 농축, DNA분절, MMP와 proteasome 활성도의 감소, Bax의 증가와 Bcl-2의 감소, DNA양의 감소, cytochrome c의 세포질로의 유리, AIF와 DFF40(CAD)의 핵으로의 이동, caspase-9, caspase-7, caspase-3, PARP 그리고 DFF45(ICAD)의 활성화와 같은 다양한 세포자멸사 증거를 보였다. 반면에 상기 물질들에 단독처리 된 SCC25 세포에서는 세포자멸사 현상이 거의 없었다. 24시간 동안 $25{\mu}M$의 HS-1200, $4{\mu}g/ml$의 cisplatin 을 각기 단독처리 한 결과에서는 세포자멸사를 거의 유도하지 못했으나, 병용처리한 결과에는 아주 탁월하고 명확한 세포자멸사의 유도를 보였다. 그러므로 본 실험결과는 HS-1200과 cisplatin 의 병용요법이 사람구강편평세포암종 환자를 위해 새로운 치료전략으로서의 가능성을 보여준다고 생각한다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제37권6호
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• pp.550-555
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• 2011

Chromosomal loss of heterozygosity (LOH) is a common mechanism for the inactivation of tumor suppressor genes in human epithelial cancers. LOH patterns can be generated through allelotyping using polymorphic microsatellite markers; however, owing to the limited number of available microsatellite markers and the requirement for large amounts of DNA, only a modest number of microsatellite markers can be screened. Hybridization to single nucleotide polymorphism (SNP) arrays using Affymetarix GeneChip Mapping 10 K 2.0 Array is an efficient method to detect genome-wide cancer LOH. We determined the presence of LOH in oral SCCs using these arrays. DNA was extracted from tissue samples obtained from 10 patients with tongue SCCs who presented at the Hospital of Tokyo Dental College. We examined the presence of LOH in 3 of the 10 patients using these arrays. At the locus that had LOH, we examined the presence of LOH using microsatellite markers. LOH analysis using Affymetarix GeneChip Mapping 10K Array showed LOH in all patients at the 1q31.1. The LOH regions were detected and demarcated by the copy number 1 with the series of three SNP probes. LOH analysis of 1q31.1 using microsatellite markers (D1S1189, D1S2151, D1S2595) showed LOH in all 10 patients (100). Our data may suggest that a putative tumor suppressor gene is located at the 1q31.1 region. Inactivation of such a gene may play a role in tongue tumorigenesis.

• Journal of Ginseng Research
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• 제17권1호
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• pp.52-60
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• 1993

Panax ginseng has been extensively used in the traditional oriental medicine as a restorative, tonic and Prophylactic agent. Recently, several reports regarding to anticancer effects of Panax ginseng has accumulated. These studies emphasized the fact that the anticancer activities might be due to a glycoside group called ginsenoside or pan.u saponin which has a water soluble characteristic. However, the authors and collaborates demonstrated that a highly lipid soluble component in extract of Panax ginseng roots contains a considerable cytotoxic activities against marine leukemic cells (L1210, P388) and human censer cells (HRT-18, HT-29, HCT48). This study was devised to observe the cytotoxic activities of Petroleum-ether extract of Panax giuseng roots (crude GBD and its Partially Purified fraction from silicic acid column chromatography (7 : 3 GX) against sarcoma-180 (5-180) and Walker carcinosar- coma 256 (Walker 256) in vivo, and murine leukemic Lymphocytes (L1210) and human rectal cancer cells (HRT-18) and human colon cancer cells (HT-29 and HCT48) in vitro. Each cell-line was cultured in medium containing serial concentration of the crude GX or 7 : 3 GX in vitro. A highly lipid soluble compound in the extract of Panax ginseng root was cytocidal to murine leukemic cells and human colon and rectal cancer cells in vitro. In the meantime, ginseng saponin derivatives did not have cytotoxic effects at its corresponding concentration. The growth rates of the cancer cells in medium containing ginseng extracts were inhibited gradually to a significant degree roughly in proportion to the increase of the extract concentration. The cytotoxic activity of 7 : 3 GX was about 3 times more potent than that of crude GX, one unit of cytotoxic activity against L1210 cells being equivalent to 2.54 Ug and 058 Ug for the crude GX and 7 : 3 GX, respectively. The Ri value of the active compound on silica- gel thin layer chromatography with petroleum-ether/ethyl ether/acetic acid mixture (90 : 10 : 1, v/v/v) as a developing so lvent was 053. While, the Panaxydol and Panaxynol as active compounds were purified from Petroleum-ether extract of Panax ginseng root by Drs. Ahn and Kim, and author found out that the one unit of cytotoxic activity of the Panaxydol and Panaxynol against L1210 cells being equivalent to 056 Ug and 0.3918 respectively. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times by the 7 : 3 GX treatment compared with their control group. The significantly decreased hemoglobin values of rats after inoculation with Walker 256 were recovered to normal range by oral administration of the crude Gt The synthetic levels of protein, DNA and RNA in human colon and rectal cancer cells were significantly diminished by treatment with the crude GX, which can explain a part of the origin of its anticancer activity.

• Journal of Nutrition and Health
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• 제23권2호
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• pp.135-147
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• 1990

본 연구는 유기용매로 추출한 마늘의 항암성 성분을 in vitro에서 흰생쥐의 백혈병성 임퐈모 세포인 L$_{1210}$과 P${388} 및 인체 직장암 세포인 HRT-18과 인체 결장암 세포인 HCT-48과 HT-29 또한 invivo에서 흰쥐 복수 육종암세포인 sarcoma-180세포를 대상으로 선별 시험한 것이다. S-180을 제외한 각 암세포는 in vitro에서 마늘추출물을 농도별로 첨가한 배양액에서 배양시험하였다. 암세포의 증식억제효과는 마늘의 석유에텔추출물이 알코올추출물보다 높았다. 마늘의 지용성 성분은 in vitro에서 흰생쥐의 백혈병성 임퐈모 세포, 인체 직장암 및 결장암 세포에 대해 항암효과물을 나타내었다. 각 암세포의 증식률은 첨가한 마늘추출물의 농도가 증가할수록 감소한 영향을 보였다. Petroleum ether/ethyl ether/acetic acid(90:10:1, v/v/v)의 전개용매로 사용한 TCL에서 분리한 유효한 활성성분의 Rf치는 0.18이었고, 정제하지 않은 마늘추출물보다 L$_{1210}$세포에 대해 2.3배 높은 활성을 나타냈다. S-180 암세포로 유발한 흰생쥐에 마늘 추출물을 투여한 군이 투여하지 않은 군보다 생존 기간이 1.5내지 2배 가량 연장효과를 보였다. S-180 암세포를 함유하고 있는 흰 생쥐에 마늘추출물(3ug/head)을 복강내 주사하고 3시간 후에 관찰한 결과 S-180 암세포의 뚜렷한 형태변화를 관찰하였다.

• 치과방사선
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• 제28권1호
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• pp.127-143
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• 1998

The author evaluated the effects of taxol, a microtubular inhibitor, as a possible radiation sensitizer and the production of prostaglandins on three human cancer cell lines(KB, RPMI-2650 and SW-13) and one murine cell line(L929). Each cell line was divided into four groups (control, taxol only, radiation only and combination of taxol and radiation). The treatment consisted of a single irradiation of 10Gy and graded doses (5, 50, 100, 200, 300, 500 nM) of taxol for a 24-h period. The cytotoxicity of taxol alone was measured at 1 day after(1-day group) and 4 days after(4-day group) the treatment. The survival ratio of cell was analyzed by MTT (3-(4,5-dimethylthiazol-2-yl) -2,5-dimethyl tetrazolium bromide) test. Prostaglandins(PGE2 and PGI2) were measured in the culture medium by a radioimmunoassay. The results obtained were as follows. 1. There was a significantly increased cytotoxicity of KB cells in 4-day group than those in I-day group. There was a high correlation between doses of taxol and cell viability in both groups(l-day group R=0.82741, 4-day group R=0.84655). 2. There was a significantly increased cytotoxicity of RPMI -2650 cells treated with high concentration of taxol in 4-day group than those in I-day group. Also there was a high correlation between doses of taxol and cell viability in 4-day group(R=0.93917). 3. There was a significantly increased cytotoxicity of SW-13 cells treated with high concentration of taxol in 4-day group than those in 1-day group. However no high correlation was observed between doses of taxol and cell viability in both groups(1-day group R=0.46362, 4-day group R=0.65425). 4. There was a significantly increased cytotoxicity of L929 cells treated with low concentration of taxol in 4-day group than those in 1-day group. At the same time, there was a low correlation between doses of taxol and cell viability in both groups(1-day group R=0.34237, 4-day group R=0.23381). 5. In I-day group of L929 cells, higher cytotoxicities were observed in the groups treated with 500 nM taxol than given 10 Gy radiation alone. L929 cells in I-day group alone showed a radiosensitizing effect by taxol.. 6. In addition to L929 cells, all cancer cells treated with a combination of taxol and radiation in 4-day group appeared to have some fragmented nuclei and to float on the medium. In addition, L929 cells appeared to be more confluent. 7. The level of PGE2 production was the highest in the contol KB cells. This appeared to increase in every experimental group of all three cancer cells except L929 cells. There was a significantly increased production of PGE2 in SW -13 cells treated with a combination taxol and radiation compared to the other experimental groups. 8. The level of PGE2 production in the control group of RPMI-Z650 cells was the highest. This appeared to increase in every experimental group of all cells except in SW-13 cells. This also increased significantly in RPMI-2650 cells treated with a combination of taxol and radiation compared to the other experimental groups.

• Journal of Oral Medicine and Pain
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• 제35권3호
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• pp.165-175
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• 2010

Valproic acid(VPA)는 아주 잘 알려진 항경련제로서, 30년 동안 간질치료제로서 사용되어져 왔다. VPA는 1997년에 최초로 원시 신경외배엽성 암세포의 증식 억제와 분화를 유도하는 항암제의 효능이 밝혀졌다. 그리고 VPA의 항암효과는 히스톤탈 아세틸화효소억제제의 기전에 기인한다고 규명되었다. 담즙산과 합성담즙산유도체가 여러 종류의 암세포에 세포자멸사(apoptosis)를 유도하며, 항암효과가 있다고 알려져 있다. 또한 합성 chenodeoxycholic acid(CDCA) 유도체가 여러 가지 암세포에 유도한 세포자멸사 연구들이 보고되어져 왔다. 본 연구는 히스톤탈아세틸화효소억제제인 VPA와 합성 CDCA 유도체인 HS-1200의 병용처리가 사람골육종세포에 효과적인 상승 세포자멸사 효과가 있는지를 알기 위해서 수행되었다. VPA과 HS-1200의 병용처리가 단독처리에 비해서 효과적인 세포생존율 감소가 있는지 확인하기 위해서 trypan-blue법을 시행하였고, 세포자멸사의 유도와 증가를 확인하기 위해서 Hoechst 염색법, flow cytometry(DNA hypoploidy와 MMP 측정), Western bot 분석법 그리고, 면역형광염색법을 수행하였다. 병용처리 된 사람골육종세포는 단독처리 된 사람골육종세포에서 거의 관찰할 수 없었던 많은 핵 응축, DNA 조각남, 사립체막 전위와 DNA 양의 감소, cytochrome c의 세포질로의 유리, AIF의 핵으로의 이동, caspase-7, caspase-3 그리고 PARP의 파괴와 같은 세포자멸사 증거를 보였다. 48시간 동안 1 mM의 VPA와 $25\;{\mu}M$ HS-1200을 각기 단독처리 한 결과에서는 세포자멸사를 유도 못했으나, 병용처리한 결과에는 아주 탁월한 세포자멸사의 유도를 보였다. 이러한 병용처리 결과는 사람골육종의 새로운 치료적 전략으로 응용될 수 있다고 생각한다.