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송이버섯(Tricholoma matsutake) 추출물이 인체 구강암 KB Cell Lines에 미치는 영향

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Author(s)
이춘식
Issued Date
2010
Keyword
Tricholoma matsutake pine mushroom Human Oral Cancer KB Cell Lines MTS Western Blotting iNOS COX-2
Abstract
이 논문은 송이버섯을 분말로 갈아서 열수와 메탄올로 각기 다른 농도의 추출물을 만들고 시간을 달리하여 저분자량 상태와 고분자량 상태에서 인체구강암 KB 세포주의 세포 생존율과 암 표적인자의 발현량을 측정하여 암 예방을 위한 보조제 및 식이요소의 가능성을 알아보고자 했다.

이 연구에서는 MTS assay 와 Western Blotting의 두 가지 실험 방법이 사용되었다. 송이버섯의 열수 추출물과 메탄올 추출물이 각각 인체구강암 KB세포주의 세포생존율에 미치는 영향을 측정하기 위하여 MTS assay를 시행했으며, 암 표적인자 COX-2와 iNOS의 발현량 억제에 미치는 영향을 측정하기 위하여 Western Blot assay를 사용하였다.

송이버섯 메탄올 추출물은 농도가 높아짐에 따라 (900 ㎍/㎖) 암 표적인자 iNOS 발현량을 놀랍게도 80%나 억제시켰고, COX-2 발현량도 40% 감소시켰다. 또한 시간이 경과함에 따라 48시간 만에 농도 900 ㎍/㎖에서 암 세포 생존율을 65%나 억제시켰다.

송이버섯 열수 추출물은 300 ㎍/㎖라는 저농도에서 암 표적인자 iNOS 발현량을 60% 억제 시켰다. 이 실험 결과는 40,000 KDa 이상의 분자량에서 300 ㎍/㎖라는 저농도를 가지고 암 표적인자 억제라는 결과를 낸 것이므로 매우 유의적이며 더 적극적으로 추가적인 실험의 필요성을 보여주는 것이라고 사료된다.

이 연구의 결론으로서, 송이버섯 추출물은 암 표적인자 iNOS 발현량의 놀라운 억제능 및 시간과 농도에 따른 암세포 생존율 억제능을 보였다. 또한 열수 추출물의 유의적 억제 효과는 저농도에서 나타났으므로 식품과정을 통한 암 예방적 가능성을 보여 준 것이라고 해석됨으로 추가적인 IN VIVO 실험이 요구된다.

또한 송이버섯에서 추출될 수 있는 약용요소 - 즉 불포화지방산, 레티난과 같은 베타 글루칸, 폴리사카라이드 K와 같은 - 에 대한 추가적인 분리 정제 실험이 필요하다고 생각된다.
The basic purpose of this study was to know whether two types of extracts from Tricholoma matsutake have or do not have the significant inhibitory effects on the cell viability of KB human oral cancer cell lines according to different molecular weights such chemoprevention as anticancer adjuvant and dietary factors.

This study was used two experimental methods of MTS assay and Western Blotting. MTS assay was performed to detect the inhibitory effects of Tricholoma matsutake extracts on the cell viability of the KB human oral cancer cell lines. Western Blotting was performed to observe the suppressive effects of Tricholoma matsutake extracts on inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression as cancer marker.

Methanol extracts from Tricholoma matsutake showed surprisingly the more than 80% suppression of iNOS expression and the 40% decrement of COX-2 expression with the progress of high consistency. Also they showed about 65% inhibitive action on the cell viability rate with the progress of high consistency as a result of treatment for 48 hours.

Hot water extracts of Tricholoma matsutake showed the significant 60% suppression of iNOS expression at the low density of the log of 300 ㎍/㎖. This result means the potentiality of chemoprevention as anticancer adjuvant therapy because it occurred in the low density and the macromolecule above 40,000 KDa molecular weight.

As the conclusion of this study, the surprising suppression of iNOS expression and cell viability presented the high possibility of chemoprevention as anticancer adjuvant and dietary factors. but researches of activating substances such as the medicinal properties from Tricholoma matsutake as unsaturated fatty acid, β-glucans like Lentinan and Polysaccharide-K etc should continue for remedies.

Also the conclusion of this study can be stated the potentiality of chemoprevention on iNOS and COX-2 in inflammatory cells by dietary factors using such as food phytochemicals and polyunsaturated fatty acid.
Alternative Title
Effects of Tricholoma matsutake Extracts on the Human Oral Cancer KB Cell Lines
Alternative Author(s)
Lee Choon-sik
Department
보건대학원 대체의학과
Advisor
문경래
Awarded Date
2011. 2
Table Of Contents
CONTENTS

LIST OF FIGURES.................................................................................. 7 page
ABSTRACT.................................................................................................. 8 page

Ⅰ. INTRODUCTION............................................................................. 10 page

Ⅱ. MATERIALS AND METHODS................................................... 14 page
1. Basic raw materials and extracting for experiments
2. Reagents .
3. Cell culture & chemical treatment
4. Preparation of Tricholoma matsutake
5. MTS assay for estimating cell viability
6. Western blot analysis (iNOS and COX-2 expression)
7. Statistical analysis

Ⅲ. RESULTS................................................................................,............... 17 page
1. Effects of Tricholoma matsutake extracts on
Human Oral Cancer KB Cell Lines viability
using MTS assay (Methanol extracts)
2. Effects of Tricholoma matsutake extracts on
Human Oral Cancer KB Cell Lines viability
using MTS assay (Hot water extracts)
3. Effects of Tricholoma matsutake methanol extracts on
COX-2 and iNOS expression (Western Blotting
in Human Oral Cancer KB Cell Lines)
4. Effects of Tricholoma matsutake hot water extracts on
COX-2 and iNOS expression (Western Blotting
in Human Oral Cancer KB Cell Lines)

Ⅳ. DISCUSSION..............................................................................,........... 22 page

Ⅴ. CONCLUSION...................................................................................... 29 page

Ⅵ. KOREAN ABSTRACT........................................................................ 31 page

Ⅶ. REFERENCES........................................................................................ 33 page
Degree
Master
Publisher
조선대학교 보건대학원
Citation
이춘식. (2010). 송이버섯(Tricholoma matsutake) 추출물이 인체 구강암 KB Cell Lines에 미치는 영향
Type
Dissertation
URI
https://oak.chosun.ac.kr/handle/2020.oak/2207
http://chosun.dcollection.net/common/orgView/200000241220
Appears in Collections:
Medicine & Parmacy > Theses(Master)(의학전문대학원/보건대학원/임상약학대학원)
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