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유방암 세포에서 황칠나무 잎 추출물의 MAPK 경로를 통한 apoptosis 유도
정기환(Gi Hwan Jung),김성현(Sung Hyun Kim),우중석(Joong Seok Woo),유은선(Eun Seon Yoo),이재한(Jae Han Lee),한소희(So Hee Han),정수현(Soo Hyun Jung),김세만(Sae Man Kim),김은지(Eun Gee Kim),신만재(Man Jae Shin),조호민(Ho Min Cho),정지윤(J 한국생명과학회 2021 생명과학회지 Vol.31 No.9
황칠나무 잎 추출물(DPL)은 염증, 당뇨병, 암 등 다양한 질병에 예로부터 약으로 사용되어 왔다. 특히 여러 종류의 암세포에 세포사멸 효과가 있는 것으로 밝혀졌지만 유방암 세포 SK-BR-3에 대한 세포사멸 효과는 아직 밝혀지지 않았다. 따라서 본 연구에서는 in vitro 실험을 통해 DPL이 유방암 세포 SK-BR-3에서의 증식 억제와 apoptosis를 통한 세포사멸 효과를 확인하였다. DPL이 세포 생존율에 미치는 영향을 조사하기 위해 MTT 분석을 수행했고 세포 생존율이 농도의존적으로 감소하는 것을 확인했다. DAPI 염색은 DPL에 의해 발생한 세포 형태학적 변화와 apoptotic body의 증가에 미치는 영향을 조사하기 위해 수행되었다. 이를 보완하기 위해 annexin V/PI 염색후 유세포 분석을 통해 세포 apoptosis 비율의 증가를 확인했다. Apoptosis 관련 단백질들을 확인하기 위해 Western blot을 수행하였으며, cleaved-PARP과 pro-apoptosis 단백질인 Bax의 증가와 anti-apoptosis 단백질인 Bcl-2의 감소를 확인하였다. MAPK 경로 단백질 p-ERK1/2, p-JNK 및 p-p38의 발현 수준의 변화도 확인하였으며 p-p38의 유의미한 증가가 관찰되었다. 본 연구 결과에 따르면 DPL은 유방암 세포 SK-BR-3에서 p-p38 MAPK 경로 조절을 통해 세포사멸 효과를 유도하는 것으로 사료된다. Dendropanax morbiferus leaves (DPL) has been used as a medicine since ancient times in various diseases such as inflammation, diabetes, and cancer. In particular, it has been found to have anticancer effects on several types of cancer cells, but the anticancer effect on breast cancer cells SK-BR-3 has not yet been revealed. Therefore, in this study, DPL caused proliferation inhibition in breast cancer cells SK-BR-3 and the anticancer effect by inducing apoptosis was confirmed, through an in vitro experiment. In order to examine the effect of DPL on cell viability, MTT assay was performed to confirm a significant decrease in the concentration of cell viability. DAPI staining was performed to examine the effect of DPL on cellular morphological changes and increase of apoptotic bodies. To supplement this, an increase in the apoptosis rate was also confirmed through flow cytometry after staining with annexin V/PI. Western blot was performed to confirm apoptosis-related proteins. DPL increased the expression of Cleaved-PARP, Bax whereas decreased the expression of Bcl-2. Changes in the expression levels of MAPK pathway proteins p-ERK1/2, p-JNK, and p-p38 were also confirmed, and a significant increase in p-p38 was observed. These results indicated that DPL induced apoptosis, through p-p38 MAPK signal pathway in SK-BR-3 breast cancer cells.
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백서에서 자가 피부이식후 항T임파구 단일클론항체 분포에 관한 면역조직화학적 연구
朴晩圭,金慶助,任昌俊,高世勳 단국대학교 치의학연구소 1991 논문집 Vol.3 No.-
This article is intended to study histopathological and immunohistochemical response after autogenous full-thickness skin graft in rat. 12 male Sprague-Dawley rats were used as the experimental animals. A 1 cm×1 cm skin (0.7mm diameter) was taken on the right inguinal area of the rat. Another full-thickness skin graft (1cm × 1cm) was taken from the left inguinal area of the rat. And it was transplanted to the right inguinal area of the rat. The left side wound was closed directly. Light microscopic observation was made at the postoperative 1, 3, 8, 16 day, after the hematoxylin-Eosin staining of the 4u-thick paraffin embedded specimens and the immunohistochemical staining of the 10u-thick frozen specimens with mouse anti-rat monoclone antibodies and ABC staining kit. The results were as follows. 1. Electromicroscopic studies revealed interstitial tissue bleeding of transplanted autogenous skin. The response was severe in the 1 day group after operation, moderate in 3 day group, mild in 8 day group, and almost resolved in the 16 days group. 2. Electromicroscopic studies also revealed a mild monocyte response in the 3 day and 8 day groups. A histiocytic infiltrate was observed. There was a mild response in the 3 day group and moderate response in the 8 day group. 3. Immunohistochemically studies revealed a few pan T cells in the 1 day group, mild appearance of pan T cells and cyototoxic T cells in the 3 day group, a moderate infiltrate of pan T cells and helper T cells in the 8 day group, and total resolution of pan T cells in the 16 day group. 4. According to these finding, a strong inflammatory response was observed around transplanted autogenous skin in the 3 & 8 day groups. In the 16 day group this response had resolved histopathologically and immunohistologically.
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