Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
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The Effect of Uteroglobin on cPLA2, COX-2 Expression and ERK Activation in NSCLC Cells

비소세포 폐암세포에서 Uteroglobin의 이입에 의한 cPLA2와 COX-2 발현 및 ERK 활성의 변화

  • Kim, Woo Jin (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Yoon, Jung Min (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Lee, Kyoung Hee (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Han, Seon Jin (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Shin, Won Hyuk (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Yim, Jae-Joon (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Yoo, Chul-Gyu (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Lee, Choon Taek (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Han, Sung Koo (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Shim, Young-Soo (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine) ;
  • Kim, Young Whan (Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine)
  • 김우진 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 윤정민 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 이경희 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 한선진 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 신원혁 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 임재준 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 유철규 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 이춘택 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 한성구 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 심영수 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소) ;
  • 김영환 (서울대학교 의과대학 내과학교실 및 의학연구원 폐연구소)
  • Published : 20040000
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Abstract

Background : Uteroglobin is a protein produced by the normal bronchial epithelium and its expression level is lower in non-small cell lung cancer tissues and cell lines. It mainly functions as an anti-inflammatory, and when it is overexpressed in cancer cells, the neoplastic phenotype is antagonized. cPLA2 and COX-2, which are also associated with inflammation, were reported to be related to cancer. The relationship between cPLA2, COX-2 and uteroglobin is unclear. The relationship between uteroglobin and ERK, which is related to cell growth, is also not unclear. This study investigated the changes in the cPLA2 and COX-2 expression levels and the ERK activities after the overexpression of uteroglobin in non-small cell lung cancer cell lines. Methods : The A549 and NCI-H460 cell lines were infected by adenovirus-null and adenovirusuteroglobin. The cChange in the cPLA2, COX-2 expression level and ERK activity after uteroglobin overexpression was measured by Western blot. The change in MMP activity was measured by zymography. Results : Western blot revealed decreased expression levels of cPLA2, and COX-2, and increased pERK levels in nonsmall cell lung cancer cells after uteroglobin overexpression. Zymography revealed no changes in the MMP-2 activity and lower MMP-9 activity. U0126, which is a specific inhibitor of ERK-activating kinase MEK-1/-2, prevented the decrease in the MMP-9 activity Conclusions : A decrease in cPLA2 expression, COX-2 expression, MMP-9 activity and a increase in ERK activity may be related to the anticancer effects of uteroglobin in nonsmall cell lung cancer cells.

연구배경 : Uteroglobin은 정상 폐상피세포에서 발현되는 단백질로 비소세포암 조직이나 세포주에서는 발현이 저하되어있다. 항염증작용을 하며 암세포에 과발현 시키면 암의 형질이 소실됨이 밝혀지고 있다. 역시 염증작용과 관련이 있는 cPLA2와 COX-2도 암과의 관련성이 밝혀지고 있고, 암억제나 MMP의 억제 등의 공통점을 가지고 있으나 이들의 관련성에 대해서는 밝혀진 바가 없다. 또한, COX-2의 암과의 관련성을 설명하는 기전으로 ERK 활성화의 관련 가능성이 있으나, uteroglobin과 ERK의 관련성도 아직 밝혀지지 않고 있다. 비소세포폐암 세포주에 uteroglobin을 과발현시킨 후, cPLA2와 COX-2의 발현, 그리고 MMP-2, MMP-9, ERK의 활성화가 어떻게 변화하는지에 대해 실험하였다. 방 법 : 폐선암세포주인 A549와 NCI-H460 세포주에 adenovirus-uteroglobin, adenovirus-null을 각각 20,100,200 moi로 transduction 시킨 뒤, 48시간 배양한 후에 단백질을 추출하였다. Uteroglobin의 발현을 확인한 후, cPLA2, COX-2, pERK, total ERK에 대해 Western blot을 시행하였고, 배양액으로 zymography를 시행하였다. 결 과 : A549 세포주와 NCI-H460 세포주에서 uteroglobin의 발현을 확인한 세포주에서 cPLA2와 COX-2의 발현이 감소함을 Western blot으로 확인하였고, pERK가 증가함을 Western blot으로 보았고, ERK의 활성화가 증가함을 확인하였다. MMP-9은 활성이 저하되었고, MMP-2는 변화가 없었다. MEK inhibitor인 U0126을 이용하여 ERK의 활성화를 저해시킨 후, uteroglobin의 발현에는 영향이 없었고, MMP-9의 활성저하효과가 소실되었다. 결 론 : 폐암세포주에서 uteroglobin의 항암작용기전에 cPLA2 와 COX-2의 발현의 감소와 ERK의 활성화가 기여할 것으로 사료된다.

Keywords

References

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