Jaejun Shim 월요저널 2010-11-22. Introduction Obesity – White adipose tissue: adipokines (leptin and adiponectin) –Leptin: apheliotrophic actions Atherosclerosis,

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1 Jaejun Shim 월요저널 2010-11-22

2 Introduction Obesity – White adipose tissue: adipokines (leptin and adiponectin) –Leptin: apheliotrophic actions Atherosclerosis, carcinogenesis Cell proliferation, migration, invasion Breast, prostate, colon cancer model

3 Introduction –Adiponectin: protective role against obesity- related disorders Glucose regulation Fatty acid catabolism Exclusively secreted from adipose tissue Inversely correlated with body fat –Obesity : inversely decreased adiponectin level Type 2 DM, obesity, atherosclorosis, NASH, metabolic syndrome Antineoplastic effect?

4 Adiponectin  leptin-induced malignant properties of HCC cells? in vitro: cell proliferation, apoptosis in vitro: molecular mechanism, effector signaling pathway – Western blotting in vivo: nude-mice Clinical correlation: tissue expression, survival and prognosis Human HCC cell lines: HepG2, Huh7 Bromodeoxyuridine (BrdU) incorporation TUNEL test Western blot Migration assay: scratch migration protocol, Cell-substrate impedance sensing (ECIS) Tumor cell invasion assay: Matrigel invasion assay HCC tumorigenesis assay Immunohistochemistry, HCC TMAN

5 Results

6 Adiponectin  apoptosis, proliferation  leptin

7 Adiponectin  migration  leptin

8 Adiponectin  invasion  leptin

9 Leptin intracellular signaling pathway JBC 2000;275:40649

10 Adiponectin  phosphorylation of leptin signaling molecule

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12 Expression of adiponectin and leptin in human HCC

13 Leptin and adiponectin expression in human HCCs

14 Leptin and adiponectin expressin in human HCCs

15 Conclusion Adiponectin has the molecular potential to inhibit the oncogenic actions of leptin by blocking downstream effector molecules

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17 Adiponectin analogue: potential therapeutics for metastatic HCC? Obesity  leptin  HCC tumorogenesis NASH  fibrosis  cirrhosis

18 Leptin Leptin: 그리스어로 thin 이라는 뜻 16kDa protein hormone, 167 amino acids 에너지 섭취와 대사에 중요한 역할을 담당 지방세포에서 분비되는 가장 중요한 호르몬임 7 번 염색체 Ob (Lep) gene 1950 년대 ob/ob mutant obese mice 발견 1994 년 Jeffrey M Friedman 록펠러 대학에서 발견 체지방과 혈중 농도와는 직접적인 상관관계 White adipose tissue 에서 생산

19 Leptin Function – 시상하부의 leptin receptor 에 작용 Neuropeptide Y 의 작용을 억제 Anandamide 억제 Alpha-MSH 식욕 억제제 생산 촉진 –Ob-Rb receptor  stat3  Jak-Stat, MAPK signal transduction

20 Adiponectin A 244-aminoacid-long polypectide Metabolic process 를 담당하는 protein hormone Glucose regulation Fatty acid catabolism Exclusively secreted from adipose tissue Inversely correlated with body fat –Obesity : inversely decreased adiponectin level Type 2 DM, obesity, atherosclorosis, NASH, metabolic syndrome

21 STAT3 Signal transducer and activator of transcription 3 also known as STAT3 is a transcription factor which in humans is encoded by the STAT3 gene.

22 pAKT

23 Electric cell-substrate impedance sensing (ECIS) Measuring changes in AC impedance of small gold-film electrodes deposited on a culture dish Cell spreading kinetics Attachment and spreading of cells

24 Matrigel Invasion Assay Protocol Matrigel is considered as basement membrane and generated from EHS sarcoma. Matrigel contains not only basement membrane components (collagens, laminin, and proteoglycans)but also matrix degrading enzymes/their inhibitors and growth factors. Invasion of tumor cells into Matrigel has been used to characterize involvement of ECM receptors and matrix degrading enzymes which play roles in tumor progression.

25 Luciferase 루시페라아제 (Luciferase) 는 루시페린을 산화시키는 효소이다. 개똥 벌레와 발광성 미생물 등에 존재하며 생물 발광에 관여하고 있다. 루 시페린은 특정한 물질을 뜻하는 말이 아니며, 발광에 관여하는 물질 을 통칭하는 명칭이다.[1]

26 Ki-67 Antigen KI-67 is a nuclear protein that is associated with and may be necessary for cellular proliferation. Furthermore it is associated with ribosomal RNA transcription The Ki-67 protein (also known as MKI67) is a cellular marker for proliferation.[5] It is strictly associated with cell proliferation. During interphase, the Ki-67 antigen can be exclusively detected within the cell nucleus