Mouse L Cell에서의 외래 유전자 유래 단백질의 생산

최윤희,岸本忠三,최차용 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.5

유전자 재조합 동물세포를 제작하여 외래 유전자 유래의 단백질을 생산하고자 그 model system으로써 부착성 동물 세포인 mouse L cell과 pBMG Neo vector, 그리고 외래 유전자로는 hNF-IL6를 선택하였다. hNF-IL6는 hIL-6 유전자의 promoter 서열에 결합하는 성질에 의해 분리된 핵단백질로서 IL-6의 전자조절 인자로 밝혀져 있다. 이 hNF-IL6는 다른 면역 관계 단백지의 전사 조절 영역에도 결합함이 밝혀져 그 중요성이 부각되고 있으나 재조합 대장균에서는 발현시 너무 빨리 분해되어 생산이 어려웠다. Plasmid내의 neomycin resistance 특성을 이용한 1차 screening, 그리고 southern blot analysis를 이용한 2차 screening에 걸쳐 hNF-IL6 발현 vector가 도입된 재조합 clone을 선택하였으며, 전사 및 발현를 확인하였고 그 발현이 MT promoter 특유의 유도기작에 따르는 것도 알 수 있었다. 생산된 hNF-IL6는 핵내에 존재함으로써 본래의 핵단백질 특성을 유지하고 있었으며 DNA binding activity도 확인되었다. 따라서 pBMG Neo vector를 사용, mouse L cell에서 발현된 외래 단백질 hNF-IL6가 본래의 활성도 유지함 대량 생산된 것을 알 수 있었고, 동물 세포에서의 외래 유전자 발현에 대한 이 system의 응용 가능성도 확인할 수 있었다. Some interleukin 6 (IL-6) transcription control factors were reported as the regulator of IL-6 expression. A nuclear protein bound to interleukin 1 (IL-1) responsive element in the IL-6 promoter region was named NF-IL-6 (nuclear factor for IL-6). This NF-IL6 was known to be very important as a transcription factor for various immuno-protein as well as for IL-6. The human NF-IL6 genes were transfected into the mouse L cells under the metallothionein promoter (MT promoter) to establish a model system for the expression of foreign gene in the mammalian cell line. Over a thousand of neomycin-resistant clones were obtained by using electroporation method, and some positive clones were screened with Southern blotting. After the selection of LBM6104 clone with high copy numbers, the expression of human NF-IL6 under the control of the MT promoter was confirmed by Northern and Western blot analysis. The MT promoter vector system became quite useful for a setup of foreign protein expression in the mammalian cell lines.

임상 ; 조기분만 시 융모양막염 유무에 따른 제대정맥혈장내 Interleukin-6, C-reactive Protein 및 지질과산화물의 변화

강명선 ( Myoung Seon Kang ),김윤하 ( Yoon Ha Kim ),김철홍 ( Cheol Hong Kim ),김기민 ( Ki Min Kim ),조문경 ( Moon Kyoung Cho ),김종운 ( Jong Woon Kim ),김조헌 ( Jo Heon Kim ),남종희 ( Jong Hee Nam ),양성렬 ( Sung Yeul Yang ),안봉환 대한주산의학회 2007 Perinatology Vol.18 No.4

목적: 조기분만 시 제대정맥혈장내 Interleukin-6(IL-6)와 C-reactive protein (CRP) 및 지질과산화물을 측정하여 조직학적 융모양막염 유무에 따른 이들의 변화와 주산기 예후에 어떤 역할을 하는가 규명하고자 하였다. 방법: 양막파열이 없는 조기분만 임부 39명(조기분만군), 만삭전 조기양막파열 임부 27명(양막파열군)에서 분만시 태반조직과 제대정맥혈을 채취하여 IL-6, CRP, 지질과산화물 농도를 측정하고 비교하였고, 태반조직의 병리학적 소견을 얻었다. 결과: 조기분만군 39예 중 8예, 양막파열군 27예 중 16예에서 조직학적 융모양막염이 있었으며, 양막파열군에서 조기분만군보다 증가하였다. 조기분만군과 양막파열군 간에 IL-6, CRP, 지질과산화물의 차이가 없었다. 조직학적 융모양막염이 있는 군에서 없는 군보다 IL-6, CRP, 지질과산화물 농도가 증가하였다. 조직학적 융모양막염이 있는 조기분만군에서 없는 조기분만군보다 IL-6, CRP, 지질과산화물 농도가 증가하였다. 조직학적 융모양막염이 있는 양막파열군에서 없는 양막파열군에 비해 CRP는 증가하였으나, IL-6와 지질과산화물 농도는 차이가 없었다. 선천성 패혈증이 의심스러웠던 태아 3명의 IL-6와 지질과산화물 농도가 증가된 소견을 보였다. 조직학적 제대염을 동반한 경우의 IL-6 농도는 동반하지 않은 경우보다 증가하였으며, 제대염이 있었던 6예 중 신생아 패혈증 의증 2명, 호흡곤란증후군 1명, 뇌실내출혈 1명으로 주산기 이환율이 높았다. 결론: 조직학적 융모양막염이 동반된 조기분만에서 제대정맥내 IL-6, CRP 및 지질과산화물이 증가하였으며, 이에 따라 주산기 이환율이 영향을 받을 것으로 사료된다. Objective: To investigate interleukin-6 (IL-6), C-reactive protein (CRP), and lipid peroxide levels in the umbilical venous plasma of preterm birth with or without histologic chorioamnionitis and to evaluate their roles in the pathophysiology in preterm labor and perinatal outcome. Methods: This cohort study included 66 cases of preterm delivery with preterm labor and intact membranes (PTL) (n=39) and preterm premature rupture of membranes (PPROM) (n=27). The umbilical venous blood samples were collected at the time of delivery. IL-6, CRP, and lipid peroxide levels were measured by ELISA Kit, latex agglutination assay, and thiobarbituric acid reaction. Histologic chorioamnionitis was diagnosed by the presence of neutrophil infiltration into the subamnionic space. Results: The prevalence of histologic chorioamnionitis was significantly higher in PPROM (59.3%, 16/27) than in PTL (20.5%, 8/39). IL-6, CRP, and lipid peroxide levels in the umbilical venous plasma of histologic chorioamnionitis were significantly higher than those without histologic chorioamnionitis. IL-6, CRP, and lipid peroxide levels in the umbilical venous plasma of PTL with histologic chorioamnionitis were significantly higher than those of PTL without histologic chorioamnionitis. CRP levels in the umbilical venous plasma of PPROM with histologic chorioamnionitis were significantly higher than those of PPROM without histologic chorioamnionitis. Three suspected neonatal sepsis patients have increased IL-6 and lipid peroxide levels in the umbilical venous plasma compared with patients without neonatal sepsis. IL-6 levels in the umbilical venous plasma of histologic funisitis were significantly higher than those without funisitis. Conclusion: Preterm birth with chorioamnionitis is associated with an increased level of IL-6, CRP, and lipid peroxide in umbilical venous blood. Preterm birth with chorioamnionitis may have an effect on perinatal outcome.

류마티스 관절염 환자의 활액과 활막조직에서의 IL - 6 의 활성도

조석구(Seok Goo CHo),이상헌(Sang Heon Lee),홍연식(Yeon Sik Hong),조철수(Chul Soo Cho),박석영(Seok Young Park),박동준(Dong Jun Park),김호연(Ho Youn Kim),이정용(Jung Young Lee),김상호(Sang Ho Kim),변광호(Kwang Ho Pyun) 대한내과학회 1993 대한내과학회지 Vol.45 No.2

Background: Interleukin-6 (IL-6), previously known as B cell stimulatory factor 2 (BSF-2) is a multifunctional cytokine that is produced by a various cells and plays an important role in the host defence mechanism such as regulation of immune response, acute phase reaction and hematopoiesis. Recent investigations have demonstrated that unregulated expression of IL-6 gene and overproducion of lL-6 were involved in the pathogenesis of cardiac myxoma, Castleman's disease, autoimmune disease, proliferative glomerulonephritis and certain lymphoid malignancies, especially plasmacytoma/multiple myelomas. We tried to evaluate the regulatory effect of IL 6 an the pathogenesis of rheumatoid arthritis (RA) which is characterized by hyperactivation of 8 cells, presence of various autoantibodies and increase in acute phase proteins and platelets. Methods: The IL-6 activity was measured by bioassay using the murine IL-6 dependent hybridoma cell lines (MH60. BSF2), ELISA method and immunohistochemical analysis. Results: 1) IL-6 activity was significantly elevated m synovial fluid from patients with rheumatoid arthritis (RA, n= 25) as compared with osteoarthrits (OA, n=6) and seronegative arthritis (SNA, n=4) (p<0.05). 2) IL-6 activity was also elevated in serum from patients with RA as compared with other disease (P <0.01). 3) High IL-6 activity and destructive changes in affected joints had a tendency to be correlated (r=0.716), And positive correlations between IL-6 activity and C-reactive proteins in serum were observed in patients with RA (r=0.752). 4) Immunohistachemical analysis demonstrated that positive-staining cells were located on the perivascular space. Conclusion: The results suggest that unregulated production of IL-6 may play an important role in the pathogenesis of RA and could explain local as well as generalized symptoms of RA. However, it is not known whether excessive production of IL-6 is a primary event in the disease process, or secondary consequence.

담관계암종에서 c-met, c-erbB-2, COX-2, IL-6의 발현과 임상 의의

주현호 ( Hyun Ho Joo ),송은영 ( Eun Young Song ),진상화 ( Sang Hwa Jin ),오상훈 ( Sang Hoon Oh ),최영길 ( Young Kil Choi ) 대한소화기학회 2007 대한소화기학회지 Vol.50 No.6

목적: 담관계암종의 형성 및 발달과정에 c-met, c-erbB-2, COX-2, IL-6 발현이 관여한다고 알려져 있으나 아직 병리적으로 분명하지 않다. 담관계암종에서 c-met, c-erbB-2, COX-2, IL-6 발현과 임상인자간의 관계를 규명하고자 한다. 대상 및 방법: 수술로 적출된 총 114예의 담관계암 조직에서 c-met, c-erbB-2, COX-2 및 IL-6 발현을 면역조직화학 방법으로 조사하였다. 상기 표지자의 발현과 상호 연관성을 분석하였다. 결과: 담관암 112예 중 34예(30.4%)에서 c-met 발현이 관찰되었고, c-erbB-2 발현은 112예 중 5예(4.5%), COX-2 과 발현은 113예 중 53예(46.9%), IL-6 발현은 113예 중 68예 (60.2%)에서 관찰되었다. 담관암이 근육외측 결체 조직까지 침윤한 경우에 c-met 발현율이 높았고(p=0.0262), 림프절 전이가 없을 때 IL-6 발현율이 높았다(p=0.0325). c-erbB-2 발현을 보이거나 COX-2 발현만을 보일 때 림프절 전이가 많았다(p=0.0442). 결론: 담관암종 발생에 c-met 유전자 이상이 일부 관여하며, 종양의 침습과 연관성이 있을 것으로 추정한다. COX-2 및 IL-6 발현도 관여할 것으로 추정하나 C-erbB-2 유전자 이상은 관여하지 않을 것으로 생각한다. 따라서, 담관암의 c-met, COX-2 및 IL-6 발현을 동시에 조사하는 것이 담관암 환자의 임상 경과를 예측하는 데 도움을 줄 것으로 생각한다. Background/Aims: c-met, c-erbB-2, interleukin (IL)-6, and cyclooxygenase (COX)-2 expressions are considered to be implicated in the carcinogenesis and progression of cholangiocarcinoma, but the molecular pathogenesis of cholangiocarcinoma is still poorly understood. We aimed to analyze the expressions of each marker and their relationships with clinicopathologic factors. Methods: One hundred and fourteen tissue samples were obtained from surgically resected specimens from patients with billiary tract cancer. The expressions of c-met, c-erbB-2, COX-2, and IL-6 were examined by immunohistochemically. The expression of each marker and correlations between these markers and clinicopathologic factors were analyzed. Results: The expression rates of each maker were as follows: c-met 34/112 (30.4%), c-erbB-2 5/112 (4.5%), COX-2 53/113 (46.9%), and IL-6 68/113 (60.2%), respectively. c-met expression was more frequently observed in cases with invasion through the adjacent connective tissues (p=0.0263). IL-6 overexpression was more frequently observed in cases with absent lymph node metastasis (p=0.0325). Either c-erbB-2 expression or COX-2 expression was significantly associated with lymph node metastasis (p=0.0442). Conclusions: The expression of c-met was closely related to the invasiveness of cholangiocarcinoma. Co-expression of c-met, COX-2 and, IL-6 showed a significant correlation with invasiveness and lymph node metastasis and these could be useful marker to guide clinical outcome in patients with cholangiocarcinoma. (Korean J Gastroenterol 2007;50:370-378)

Provisional Designation of the Interleukin-6 Receptor (IL-6R) as a Novel Marker Gene for Exercise Tolerance

Richard Webb(Richard Webb ),Alison Early(Alison Early ),Bethan Scarlett(Bethan Scarlett ),Jack Andrew Clark(Jack Andrew Clark ),Jumo Doran(Jumo Doran ),Daniel Nash(Daniel Nash ),Michael G Hughes(Micha 사피엔시아 2021 Exercise Medicine Vol.5 No.-

Objectives: Genomic markers linked to exercise-associated health benefits and/or sporting performance are increasingly used to guide decision-making in healthcare and sport/exercise science. This project investigated whether the IL-6R SNP “rs2228145” might be provisionally designated a novel physical activity/exercise marker. rs2228145 results in an Aspartate358/Alanine358 change adjacent to the site where the IL-6R protein is cleaved into two fragments, resulting in ~two-fold increases in blood-borne levels of soluble IL-6R [‘sIL-6R’]. Methods: Cohorts of staff/students at Cardiff Metropolitan University donated/completed: [i] finger-prick capillary blood samples (subjected to ELISAs for sIL-6R, the associated signalling protein sgp130, and the IL-6/sIL-6R complex); [ii] cheek-swab samples containing buccal epithelial cell DNA (subjected to PCR-based IL-6R/rs2228145 genotyping assays); [iii] International Physical Activity Questionnaires (to estimate physical activity levels in the week preceding sample donation). Results: As expected, we observed significant genotype-dependent differences in blood-borne sIL-6R levels (CC (44.1±21.7ng/mL) vs. AC (28.6±7.3ng/mL) vs. AA (19.9±6.5ng/mL; P<0.05)); Importantly, AA homozygotes undertook significantly more physical activity than AC heterozygotes (6318±899 v. 3904±2280 MET-mins/week; P<0.01). Genotype was significantly associated with physical activity levels (P<0.05), and sIL-6R (P=0.197) and sgp130 (P=0.160) showed non-significant correlations with physical activity levels. Conclusions: These data suggest that IL-6R/rs2228145 genotype may influence participation in physical activity/exercise, perhaps by impacting on abilities to tolerate activity without experiencing adverse-effects. Although more research is required to confirm these preliminary findings, designation of IL-6R/rs2228145 as a novel marker, and determination of participants’ IL-6R/rs2228145 genotypes, may in future be useful tools to aid exercise-providers in designing personalised exercise programmes matched to clients/patients.

복합운동 수행이 비만청소년의 혈중 C-reactive protein, Interleukin-6, 혈중 지질 및 인슐린 저항성에 미치는 영향

김정규 ( Jung Kyu Kim ),신영오 ( Young Oh Shin ),문희원 ( Hee Won Moon ) 한국스포츠정책과학원(구 한국스포츠개발원) 2007 체육과학연구 Vol.18 No.2

본 연구에서는 비만청소년을 대상으로 저항성운동과 유산소운동을 병행한 복합운동을 수행하는 것이 혈중 C-reactive protein(CRP), interleukin-6(IL-6), 혈중 지질 및 인슐린 저항성(HOMA-IR)에 미치는 영향을 알아보고자 하였다. 연구 대상은 체지방률 30% 이상의 비만청소년(n=10)이었고 저항성운동(60% 1RM)과 유산소운동(60% HRR)으로 구성된 복합운동을 주 4회 8주 동안 수행하도록 하였다. 비만청소년의 혈중 CRP와 IL-6 및 인슐린 저항성 지표를 성별과 연령이 일치하는 정상청소년(n=10)의 변인들과 비교하였고, 복합운동 수행이 이들 변인에 미치는 영향을 알아보기 위해 운동 전후의 수치들을 비교하였다. 연구 결과, ① 복합운동 수행은 비만에서 증가되었던 체중, BMI, 체지방률 및 체중에 대한 제지방량의 비율을 유의하게 감소시켰다. ② 혈중 지질 성분인 TC, TG, LDL-C, HDL-C 수치는 복합운동 수행 후 유의하게 변화되지 않았으나 대조군에 비해 유의하게 높았던 인슐린 농도와 HOMA-IR 지표는 8주 동안의 복합운동 수행 후 유의하게 감소하였다. ③ 혈중 CRP와 IL-6가 대조군에 비해 비만청소년에서 유의하게 증가되어 있었으나 복합운동 수행 후 유의하게 감소된 것을 확인하였다. 결론적으로, 비만청소년을 대상으로 저항성운동과 유산소운동을 병행한 복합운동을 수행하는 것은 비만으로 인한 전신적 염증반응을 유의하게 억제함을 혈중 CRP와 IL-6가 감소된 것으로 알 수 있었고, 이들과 밀접한 상관성이 있는 인슐린 저항성 또한 감소시킨다는 것을 확인하였다. The purpose of this study is to assess the effects of combined aerobic and resistance exercise (aerobic exercise at 60% HRR, resistance exercise at 60% 1RM) on plasma C reactive-protein (CRP), interleukin-6 (IL-6), lipids, and insulin resistance (HOMA-IR) in obese adolescent. Exercise program was performed for 8 weeks. Body composition and blood biochemical variables were analyzed before and after exercise training. In the results, there were significant differences between obese adolescent group and age-matched control group. Body weight, %fat, BMI, plasma insulin, HOMA-IR, CRP, and IL-6 in obese adolescent were increased significantly than in control group. However, these variables including plasma CRP, IL-6, and HOMA-IR in obese adolescent were decreased significantly after exercise. In conclusion, combined aerobic and resistance exercise program in obese adolescent appears to be helpful for improving the obesity and lowering obese-induced systemic inflammation.

Glycated Serum Albumin Induces Interleukin-6 Expression in Vascular Smooth Muscle Cells

Seung-Il Baek(백승일),Byung-Yong Rhim(임병용),Koanhoi Kim(김관회) 한국생명과학회 2011 생명과학회지 Vol.21 No.1

Glycate화된 단백질이 혈관질환의 발생에 관여하는지 알아보기 위하여 glycated albumin (GA)이 혈관평활근세포에서 인터루킨-6 발현에 영향을 주는지 조사하고 또한 그 기전을 구명하였다. GA에 노출된 혈관평활근세포에서 인터루킨-6 transcript가 증가하고, 인터루킨-6 단백질의 분비가 증가하고, 또한 인터루킨-6 유전자의 promoter가 활성화되었다. GA에 의한 인터루킨-6 유전자의 promoter 활성화는 dominant negative 형태의 Toll-like receptor (TLR)-4와 myeloid differentiation factor 88 (Myd88)에 의하여 크게 감소되었지만, dominant negative 형태의 TLR-2와 TIR-domain-containing adapter-inducing interferon-β (TRIF)의 영향을 받지 않았다. 그리고 Extrcellular signal-related kinase (ERK) 억제 물질들은 GA에 의한 인터루킨-6의 분비 및 인터루킨-6 유전자 promoter 활성화를 억제하였다. 그리고 인터루킨-6 유전자의 promoter의 NF-κB-binding sequence에 변이는 GA에 의한 인터루킨-6 유전자의 promoter 활성화 억제하였다. 이러한 결과는 혈관평활근세포에서 GA에 의한 인터루킨-6 유전자 활성화에 TLR-4와 ERK 및 NF-κB가 관여함을 의미한다. Diabetes mellitus is associated with vascular complications. Diabetic patients exhibit high levels of glycated adducts in serum compared to non-diabetic individuals. The aim of this study was to investigate whether extracellular glycated albumin (GA) predisposes vascular smooth muscle cells (VSMCs) to pro-inflammatory phenotype. Exposure of rat aortic smooth muscle cells (AoSMCs) to GA not only enhanced interleukin-6 (IL-6) release but also activated promoter activity of the IL-6 gene. GA-induced IL-6 promoter activation was suppressed by dominant-negative forms of Toll-like receptor (TLR)-4 and myeloid differentiation factor 88 (MyD88), but not by dominant-negative-forms of TLR-2 and TIR-domain-containing adapter-inducing interferon-β (TRIF). Extracellular signal-regulated kinase (ERK) inhibition and diphenyleneiodium (DPI) also attenuated IL-6 induction by GA. Mutation at the nuclear factor-κB (NF- κB)-binding site in the IL-6 promoter region suppressed promoter activation in response to GA. The present study proposes that GA would contribute to inflammatory reaction in the stressed vasculature by inducing IL-6 in VSMCs, and that TLR-4, EKR, and NF-κB play active roles in the process.

식이유도 비만 생쥐의 식이제한과 운동처치 후 대식세포와 단핵구의 IL-6 및 TNF-α생성의 변화

김기진 ( Ki Jin Kim ) 한국운동생리학회(구-한국운동과학회) 2010 운동과학 Vol.19 No.2

본 연구는 6주간 식이조절에 의해서 유발된 비만상태의 수컷 생쥐(C57Bl/6)를 대상으로 8주간에 걸친 탄수화물 섭취량 변화를 중심으로 한 식이제한 및 트레드밀을 이용한 운동요법을 적용하여 면역기능과 관련된 혈액세포변인의 변화와 함께 리포다당복합체 자극에 대한 복강내 대식세포의 IL-6 및 TNF-α 분비능력을 분석하여 비만상태에 의해서 저하된 면역기능 개선효과를 비교한 결과는 다음과 같다. 첫째, 식이제한 혹은 운동프로그램을 적용하여 비만처치를 실시한 그룹은 모두 현저하게 감소된 체중을 나타냄으로써 처치효과가 일정하게 나타났으며, 특히 식이제한 그룹이 현저한 체중감소를 나타냄으로서 체중감소에는 식이제한이 가장 효과적인 것으로 간주되었다. 이에 반해서 운동그룹과 식이제한 및 운동처치를 병행한 그룹은 비만유도 후 일반식이 통제그룹보다 낮은 체중을 나타냈으나 유의한 체중차이를 나타내지 않음으로서 운동프로그램의 적용이 체중감소효과에는 식이제한방법보다 상대적으로 미흡한 것으로 간주되었다. 둘째, 식이처치에 의한 비만유도의 생쥐에서 과도한 지방섭취에 의한 비만상태는 말초혈액의 T-임파구를 중심으로 한 면역관련 세포의 감소를 초래하며 비만처치에 의한 체중감소는 말초혈액의 임파구 및 T-임파구 증가에 도움을 줄 수 있다고 생각된다. 셋째, 과도한 지방식이 섭취에 의한 비만유도는 대식세포에서의 LPS 자극에 대한 사이토카인 분비능력이 저하되며, 식이제한 및 운동프로그램 적용에 의한 비만처치 시 대식세포에서 LPS 자극에 대한 TNF-α 및 IL-6의 분비량이 현저히 증가하여 면역기능의 개선 가능성이 높은 것으로 간주되었다. This study was performed the analysis of blood cell numbers related immune function and lipopolysaccharide (LPS) stimulated production of tumor necrosis factor (TNF)-α and interleukin (IL)-6 by retroperitoneal macrophage in diet-induced obesity and obesity intervention of male C57Bl/6 mice. Diet-induced obesity was fed a high-fat or a standard chow diet for 6 weeks, and obesity intervention was drived by treadmill running exercise program or food intake restriction. Test items were measured by blood cell numbers related immune function and LPS stimulated production of TNF-α and IL-6 by retroperitoneal macrophage. Subjects of diet-induced obesity showed a significant increase of body weight as compared to normal diet group. High fat diet (HFD)-diet restriction (DR), HFD-exercise (EX) and HFD-DR-EX groups showed a significant decrease of body weight after 8 week intervention as compared to HFD-HFD group. And HFD-DR showed the highest decrease of body weight among intervention groups. Diet-induced obese groups showed a decreasing tendency of blood cell numbers related immune function, and the intervention groups showed an increase of blood cell numbers related immune function after 8 week intervention. LPS stimulated production of TNF-α by retroperitoneal macrophage in diet-induced obesity showed a significant decrease than a ND group, and LPS stimulated production of TNF-α and IL-6 by retroperitoneal macrophage in intervention groups showed a significant decrease after 8 week of food intake restriction or exercise training. In conclusion, diet-induced obesity showed a attenuation of immune function, and improved the blood cell numbers related immune function and LPS stimulated production of TNF-α and IL-6 by retroperitoneal macrophage after exercise intervention and food intake restriction.

Interleukin-4 Induces Senescence in Human Renal Carcinoma Cell Lines through STAT6 and p38 MAPK

Kim, Hag Dong,Yu, Su-Jin,Kim, Hee Suk,Kim, Yong-Jin,Choe, Jeong Min,Park, Yun Gyu,Kim, Joon,Sohn, Jeongwon American Society for Biochemistry and Molecular Bi 2013 The Journal of biological chemistry Vol.288 No.40

<P>Interleukin (IL)-4, originally identified as a lymphocyte growth factor, can directly inhibit growth of certain tumor cell types. We reported previously that IL-4 induced cell cycle arrest in G<SUB>1</SUB> phase through an increase in p21<SUP>WAF1/CIP1</SUP> expression in human renal cell carcinoma (RCC) cell lines. In the present study, we investigated the underlying mechanism of IL-4-induced growth inhibition. In four of six human RCC cell lines, including Caki-1, A498, SNU482, and SNU228, IL-4 induced cellular senescence as demonstrated by enlarged and flattened morphology, increased granularity, and senescence-associated-β-galactosidase (SA-β-gal) staining. Signal tranducer and activator of transcription 6 (STAT6) and p38 MAPK were found to mediate IL-4-induced growth inhibition and cellular senescence. Both of these molecules were activated by 10 min after IL-4 treatment, and inhibition of their activity or expression prevented growth suppression and cellular senescence induced by IL-4. Inhibiting or silencing either STAT6 or p38 MAPK alone partially reduced the effect of IL-4, whereas inhibiting or silencing both molecules exerted an additive effect and almost completely abrogated the effect of IL-4. Thus STAT6 and p38 MAPK appeared to independently mediate IL-4-induced growth inhibition and cellular senescence. The p21<SUP>WAF1/CIP1</SUP> up-regulation that accompanied growth inhibition and cellular senescence by IL-4 was also attenuated additively when p38 MAPK and STAT6 were silenced. Taken together, these results show that IL-4 induces cellular senescence through independent signaling pathways involving STAT6 and p38 MAPK in some human RCC cell lines.</P>

Role of Interleukin(IL)-6 in NK Activity to Hypoxic-Induced Highly Invasive Hepatocellular Carcinoma(HCC) Cells

( Hwan Hee Lee ),( Hyojung Kang ),( Hyosun Cho ) 한국미생물생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.7

Natural killer (NK) cell dysfunctions against hepatocellular carcinoma (HCC) in a hypoxic environment. Many solid tumors are present in a hypoxic condition, which changes the effector function of various immune cells. The transcription of hypoxic-inducible factors (HIFs) in cancer cells make it possible to adapt to their hypoxic environment and to escape the immune surveillance of NK cells. Recently, the correlation between the transcription of HIF-1α and pro-inflammatory cytokines has been reported. Interleukin (IL)-6 is higher in cancers with a highly invasive ability, and is closely related to the metastasis of cancers. This study showed that the expression of HIF-1α in HCC cells was associated with the presence of IL-6 in the environment of HCC-NK cells. Blocking of IL-6 by antibody in the HCC-NK interaction changed the production of several cytokines including TGF-β, IL-1, IL-18 and IL-21. Interestingly, in a co-culture of HIF-1α-expressed HCC cells and NK cells, blocking of IL-6 increased the production of IL-21 in their supernatants. In addition, the absence of IL-6 significantly enhanced the cytotoxic ability and the expression of the activating receptors (NKG2D, NKp44, and NKG2C) in NK cells to HIF-1α-expressed HCC cells. These effects might be made by the decreased expression of HIF-1α in HCC cells through the inhibited phosphorylation of STAT3. In conclusion, the absence of IL- 6 in the interaction of HIF-1α-expressed HCC cells and NK cells could enhance the antitumor activity of NK cells to HCC cells.