구강편평상피세포암종에서 CGM과 독소루비신 병용 처리에 의한 세포자멸사 유도와 전이억제 효과

강해미,ZUNDUIJAMTS ODGEREL,김인령,박봉수 대한구강해부학회 2021 대한구강해부학회지 Vol.42 No.1

구강암은 두경부 영역에 발생하는 악성종양으로 국소적 침윤과 경부 림프절 전이가 빈번하여 사망률이높고 전 세계적으로 발생 빈도가 증가하고 있는 추세이다. 외과적 수술, 방사선요법, 항암제를 통한 다각적인 치료에도 불구하고 환자의 5년 생존율이 약 50% 정도밖에 되지 않으며 치료에 사용되는 항암제는 다양한 부작용을 동반하는 경우가 많으므로 기존 약물의 부작용을 최소화 하면서 동등한 효력을 가지는 항암제의 개발이 요구되는 실정이다. 본 연구에서는 Pistacia lenticulus 나무의 잎과 줄기에서 추출한 천연 추출물인 CGM과 기존 항암제인 독소루비신을 병용 처리하여 세포자멸사를 유도하였으며 암세포의 이동력과 침습력을 억제 시켰다. 또한 암의 전이에 관여하는 EMT 관련 단백질의 발현을 확인하였다. 그 결과 세포자멸사의 내인성 경로를 통하여 세포사멸사가 유도되었으며, EMT관련 단백질인 E-cadherin의 발현이 증가하였으며 N-cadherin, snail. slug의 발현이 억제되는 것을 확인할 수 있었다. 따라서 본 연구에서는 CGM과독소루비신의 병용 처리를 통하여 CGM단독처리보다 낮은 농도로 효과적인 항암 활성 효과를 유도하였으며, CGM이 구강암의 치료제로서 효용가능성이 있음을 밝혔다.

구강편평상피세포암종에서 Elaiophylin이 유도하는 세포자멸사와 상피간엽이행 억제 효과

강해미,박시찬,박봉수,김인령 대한구강해부학회 2023 대한구강해부학회지 Vol.44 No.1

Elaiophylin은 Streptomyces melanosporus 균에서 분리한 화합물로 항균, 항바이러스, 면역억제 등의 약리효과를 나타내며 발표된 논문에 따르면 난소암, 폐암, 흑색종 등 다양한 종양세포에서 항암활성 효과를나타내는 것으로 입증된 바 있다. 그러나 구강편평상피세포암종에서의 elaiophylin의 약리효과에 대해서는 아직 밝혀진 바가 없으므로 본 연구에서는 구강편평상피세포암종 세포주인 Ca9-22와 CAL-27세포에elaiophylin을 처리하여 세포자멸사와 상피간엽이행 매커니즘에 미치는 영향을 확인함으로써 elaiophylin이구강암의 치료제로서의 효용가치가 있는지 확인하기 위한 실험을 시행하였다. 그 결과 elaiophylin은 Ca9- 22와 CAL-27 세포에서 내인성 경로를 통한 세포자멸사를 유도하였고 암세포의 상피간엽이행에 관여하는단백질의 발현을 조절하여 항암 활성 효과를 나타내었음을 입증하였다. Elaiophylin, isolated from Streptomyces melanosporus, exhibits pharmacological effects, such as antimicrobial, antiviral, and immunosuppressive activities. According to published studies, elaiophylin has been proven to demonstrate anticancer activity in various tumor cells including ovarian cancer, lung cancer, and melanoma. However, the pharmacological effects of elaiophylin on oral squamous cell carcinoma (OSCC) cells have not been elucidated. Therefore, in this study, we treated the OSCC cell lines Ca9-22 and CAL-27 with elaiophylin to investigate its impact on cell apoptosis and the epithelial-mesenchymal transition (EMT) mechanism. The study aimed to assess the potential of elaiophylin as a therapeutic agent for oral cancer. The results revealed that elaiophylin induced apoptosis through intrinsic pathways in Ca9-22 and CAL-27 cells and demonstrated anticancer activity by regulating the expression of proteins involved in the epithelial-mesenchymal transition. These findings support the potential of elaiophylin as a treatment for oral cancer.

사람 구강암세포에서 쿼세틴에 의한 항암 및 항전이 효과에 관한 융합연구

강해미,길종진,박봉수,강현경,김인령 한국융합학회 2018 한국융합학회논문지 Vol.9 No.10

본 연구는 천연 플라보노이드계 물질인 쿼세틴을 사람 구강암세포주인 HSC-2 세포에 처리한 후 나타나는 항암 및 항전이 효과를 보기 위함이다. 연구 결과를 통하여 쿼세틴은 HSC-2 세포의 세포생존율 과 세포증식율이 감소하였으며, 이 세포 사멸의 경로는 세포 자살 프로그램인 세포자멸사의 경로를 통하여 일어나는 것임을 확인하였다. 또한 세포사멸에 큰 영향을 주지 않았던 농도인 쿼세틴 100 μM 처리군부터 HSC-2 세포의 이동력 과 침습력을 억제하는 효과가 있음을 확인 하였다. 따라서 본 연구자는 종합적인 실험 결과를 통하여 구강암 세포에 있어서 200 μM이상의 쿼세틴 처리는 세포자살 프로그램을 가동하여 세포사멸을 유도하고, 100 μM 이상의 쿼세틴 처리는 세포의 이동과 침습을 억제하여 항암, 항전이 활성을 일으키는 것을 밝혔다. 그러므로 쿼세틴은 구강암에 있어 전이를 억제하고 암을 치료하기 위한 항암제로서 충분한 가치가 있음을 시사한다. This study was conducted to investigate the anticancer and antitumor effects of quercetin, which is a natural flavonoid substance in human oral cancer cell line HSC-2 cells. The results of this study showed that quercetin reduses the cell viability and the cell proliferation rate, and it led to the evidences of cell death through apoptosis pathway. Also, lower concetration quercetin over 100 μM were inhibited the cell migration and invasion. In the present study, we conclude that quercetin treatment of more than 200 μM induces apoptosis by activating programed cell death and quercetin treatment of 100 μM or more inhibits the cell migration and invasion rate in oral cancer cells. Therefore, this study suggests that quercetin is of sufficient value as an anticancer drug to inhibit metastasis and to treat cancer.

티보자닙 처리에 의한 OSCC 세포의 항암활성 효과

강해미,이종원,김인령,박봉수 대한구강해부학회 2023 대한구강해부학회지 Vol.44 No.1

티보자닙(Tivozanib)은 주로 신세포암 치료에 사용되는 티로신 키나아제 억제제(Tyrosine Kinase Inhibitors, TKIs)로 혈관 내피 성장인자 수용체(Vascular Endothelial Growth Factor Receptors, VEGFRs) 를 표적으로 하여 종양으로 가는 혈액 공급을 억제함으로써 암세포의 성장과 전이를 억제하는 것으로 알려져 있다. 신장에 발생하는 종양과 티보자닙의 연관성에 관한 논문은 비교적 많은 보고가 된 바 있으나 구강암에서의 티보자닙의 항암활성 효과는 아직 밝혀진 바가 거의 없다. 따라서 본 연구에서는 구강편평상피세포암종 세포주인 Ca9-22와 CAL-27세포에 티보자닙을 처리하여 암 발생 과정과 밀접한 관련이 있는 세포자멸사와 자가포식작용 사이의 연관성을 밝힘으로써 구강암의 치료제로서의 효능을 입증하기 위하여 시행되었다. 실험 결과 티보자닙은 Ca9-22와 CAL-27 세포에서 내인성 경로를 통한 세포자멸사와 초기 자가포식 작용 연관 단백질의 발현을 유의미하게 조절함으로써 효과적으로 항암 활성 효과를 나타내었음을 입증하였다. Tivozanib is a tyrosine kinase inhibitor (TKI) used in the treatment of renal cell carcinoma. It targets vascular endothelial growth factor receptors (VEGFRs) within the tumor, inhibiting the blood supply to the tumor, which in turn, suppresses the growth and spread of cancer cells. While there have been numerous reports highlighting the association between diseases in the kidneys and Tivozanib, its pharmacological effects on oral cancer remain largely unexplored. Therefore, this study aimed to investigate the relationship between Tivozanib and the mechanisms related to cancer cell death (apoptosis) and autophagy in Ca9-22 and CAL-27 oral squamous cell carcinoma cell lines. The objective was to demonstrate an increased efficacy in treating oral cancer cells. The results showed that Tivozanib effectively exhibited its anti-cancer activity by regulating the expression of proteins associated with intrinsic cell apoptotic pathways and early autophagy in Ca9-22 and CAL-27 cells.

OSCC 세포에서 코디세핀이 유도하는 세포자멸사와 EMT 억제효과

신현철,강해미,박봉수,준두이젬츠 오제렐,김인령 한국구강보건과학회 2022 한국구강보건과학회지 Vol.10 No.2

Objectives: Oral squamous cell carcinoma (OSCC) is an extremely malignant cancer and invades the surrounding tissue in the head and neck region. Cordycepin, also known as 3-deoxyadenosine, is a major compound and has been suggested to have anticancer potential. In this study, we examined the anti-cancer potential of cordycepin as well as its effects on the apoptosis and EMT in Ca9-22 and CAL-27 cells. Methods: The cytotoxicity effects of cordycepin were assessed using an MTT assay. The cell morphology and mitochondrial membrane potential (MMP) changes were monitored using Hoechst and JC-1 dye. Cell cycle arrest was detected using a FACS instrument and the expression of protein level was using a western blot analysis. Results: Cordycepin treated group was significantly induced apoptosis and cell cycle arrest. Also, cordycepin inhibits the EMT in Ca9-22 and CAL-27 cells. Conclusions: Therefore, these results suggest that cordycepin treatment of OSCC cells induced anti-cancer activity.

Mechanism underlying Chios gum mastic-induced cell cycle arrest and apoptosis of the G361 human melanoma cell line

김인령,강해미,유수빈,박봉수 대한구강해부학회 2017 대한구강해부학회지 Vol.38 No.1

Chios gum mastic (CGM) is a resin extracted from the stem and leaves of Pistacia lentiscus L. var. chia. It has been used as a traditional medicine in many Mediterranean countries. Recently, numerous researches have demonstrated that CGM induces cell cycle arrest and apoptosis in many cancer cells. In the present study, an alteration of the cell cycle and induction of apoptosis by CGM treatment on malignant melanoma was investigated. CGM treatment showed the inhibition of cell viability in a dose- and time-dependent manner on the G631 melanoma cell line. Apoptotic hallmarks, such as nuclear condensation and DNA fragmentation, were also identified in CGM-treated cells. Several lines of apoptotic manifestation were demonstrated. The proapoptotic factor Bax was increased in a time-dependent manner, leading to MMP loss, proteasome activity reduction, AIF translocation, and cytochrome c release. Activation of caspases, such as caspase-9, -7, and -3, led to the cleavage of PARP and DFF45 (ICAD). DFF40 (CAD) was translocated into the nucleus from the cytoplasm of the CGM-treated G361 cells. CGM halted the cell cycle progression by G1 arrest and the reduction in the level of cyclin-dependent kinases (CDKs). Accumulation of p53 protein was not observed in cells treated with CGM for 24–72 h. The level of p27KIP1 was increased by CGM treatment only for an initial 24-h period. Cyclin D1, D3, and E protein levels were diminished in CGM-treated cells. Therefore, it is possible that CGM treatment could serve as a novel therapeutic strategy against human melanoma.

Gartanin Induces Apoptosis in Oral Squamous Cell Carcinoma Cell Lines through Mitochondrial Signaling Pathway

김인령,유수빈,강해미,박봉수 대한구강해부학회 2015 대한구강해부학회지 Vol.36 No.1

Apoptosis is a physiological process of the controlled elimination of unhealthy or damaged cells1). When cells were undergoing apoptosis, it would be show nuclear condensation, DNA fragmentation, membrane blebbing through various signaling pathway 2, 3). Mitochondria signaling pathway is among the apoptosis pathway. Mitochondria play an important role in the regulation of cell apoptosis. Changes in the mitochondria membrane potential (MMP) are considered an early event in apoptosis and many pro-apoptotic proteins can be released from the mitochondria into the cytoplasm when the MMP is damaged4). In recent studies, reported that apoptosis might play an important role in various diseases. Especially of cancer is closely associated with apoptosis. So, it is important to study the anti-cancer capacities of natural compounds for the development of anticancer drugs without side effects5). Oral cancer is the most common type of human cancers in worldwide6), and oral squamous cell carcinoma (OSCC) arises from the mucosa of oral cavity frequently7, 8). The major risk factors of oral cancer are tobacco, alcohol and high risk Human Papilloma Virus (HPV) infection. The overall survival rate has not changed in recent years, despite extensive research on the biological and molecular aspects of OSCC9). Many tropical fruits have interesting molecular biological activities with potential treatment applications. Garcinia mangostana (mangosteen) has a long history of usage as a medicinal plant in southeast asia10, 11). This tropical plant is famous for its nutritious and flavorful fruits. The pericarps of Garcinia mangostana contain numerous xanthones with chemical diversity12). Several studies have shown that typical magosteen products, α-mangostin, β-mangostin, γ-mangostin, and gartanin have various effects in diseases such as anti-oxidant, anti-inflammatory, anti-allergic, anti-bacterial, anti-cancer via cell cycle arrest and apoptosis10, 11, 13, 14). Although recently mangosteen products, research has been actively promoted but effect in the oral cavity cancer, there is insufficient. In this study, we evaluated the anti-cancer effects of gartanin, one of the major xanthones from the mangosteen fruit in OSCC cell lines.

구강편평상피세포암종에서 shikonin에 의한 상피간엽이행 억제

박단비,박봉수,강해미,유수빈,김인령 대한구강해부학회 2019 대한구강해부학회지 Vol.40 No.1

구강암은 주변 근육이나 골조직으로의 침습과 경부 림프절로 전이가 빈번히 일어나며, 전이가 일어난 환자의 5년 생존율은 약 40%로, 환자의 예후 개선을 위해 전이를 억제하는 항암제의 개발이 필요한 실정이다. 상피간엽이행 (EMT)이 일어난 세포에서는 이동성과 침습성이 증가하고, 암세포가 새로운 기관으로 전이가 일어나게 된다. 따라서, EMT를 억제하는 것은 암세포의 전이를 억제하는 치료 방법으로 고려 되고 있다. Shikonin은 지치에서 분리 정제된 물질로 항염증 효과가 널리 알려져 있다. 최근 shikonin은 다양한 암세포에서 세포자멸사 및 세포주기정지를 유도하고, 폐 및 위암세포의 전이성을 상실시키는 것이 밝혀졌다. 하지만 여전히 shikonin이 구강암에서 항전이 효과는 아직 불분명하다. 따라서 본 연구에서는 shikonin이 처리된 구강암 세포주에 대한 EMT 억제의 억제 효과를 확인 하였다. 본 연구에서 사용된 구강암 세포주 CAL-27로 세포생존율 및 세포증식율, 세포이동 및 침습, 단백질 및유전자의 발현을 확인하였다. CAL-27 세포는 shikonin 처리에 의해 생존율 및 증식율이 억제되었다. 또한, shikonin은 wound-healing 및 transwell invasion assay를 통해 CAL-27 세포의 이동 및 침습을 억제 하는것을 확인 하였다. Shikonin은 CAL-27 세포에서 epithelial marker 인 E-cadherin 의 protein 발현을 증가시켰으며, mesenchymal marker 인 N-cadherin, Snail 및 Slug 의 protein 및 RNA 발현을 감소 시켰다. 따라서, shikonin은 구강암세포주인 CAL-27의 상피간엽이행을 억제 함을 알 수 있었으며, 본 연구 결과는shikonin이 항전이성을 가지는 항암제로서의 가능성을 시사한다.

Ursolic acid induced apoptosis in human osteosarcoma cell lines

최인수,김영진,강해미,준드이젬츠 오제렐,김인령,박봉수 대한구강해부학회 2022 대한구강해부학회지 Vol.43 No.1

Ursolic acid is a polyphenolic component present in large amounts in apple peels. It possesses antioxidant, anti-fungal, and anti-inflammatory properties and inhibits the growth and metastasis of various cancer cells. However, studies on the effect of ursolic acid in osteosarcoma cells remain limited. Therefore, we investigated the role of ursolic acid-induced apoptosis and autophagy in HOS and U2OS osteosarcoma cells. Ursolic acid decreased cell survival and proliferation, and evidence of apoptosis, such as nuclear condensation and cleavage, was observed in both cell lines. Ursolic acid also significantly activated the expression of Atg5 and LC3B proteins involved in autophagy. The combination of 3-MA and ursolic acid, an autophagy inhibitor, significantly increased the expression of proteins involved in apoptosis. Therefore, ursolic acid effectively induced apoptosis and autopoiesis in osteosarcoma cells and was more effective when treated with an autopoiesis inhibitor.

HS-1200 Induces Apoptosis and Inhibits Epithelial-Mesenchymal Transition in Oral Squamous Carcinoma

김영진,최인수,강해미,준두이젬츠 오제렐,김인령,박봉수 한국구강보건과학회 2022 한국구강보건과학회지 Vol.10 No.4

Objectives: HS-1200 is a synthetic derivative of bile acid that has been proven to induce apoptosis in various types of cancer cells. However, whether HS-1200 inhibits metastasis in oral cancer has not yet been determined. Therefore, this study aims to evaluate the anti-cancer effect of HS-1200 through the inhibition of EMT in human oral squamous cell carcinoma (OSCC). Methods: The cytotoxicity effect of HS-1200 was assessed using an MTT assay. Furthermore, the cell migration and invasion ratios were obtained using a wound-healing assay and a transwell migration assay. The expressions of protein and mRNA levels were measured using a western blot analysis and real-time PCR. Results: HS-1200 was observed to inhibit the metastasis of cancer cells by regulating EMT in SCC-25 and HSC-3 cells. Conclusions: HS-1200 exhibits considerable potential as a treatment for OSCC.