中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2013年
7期
524-527
,共4页
周莹%张博恒%殷欣%任正刚
週瑩%張博恆%慇訢%任正剛
주형%장박항%은흔%임정강
癌,肝细胞%趋化因子,CCL28%缺氧诱导因子-1α%侵袭
癌,肝細胞%趨化因子,CCL28%缺氧誘導因子-1α%侵襲
암,간세포%추화인자,CCL28%결양유도인자-1α%침습
Carcinoma,hepatocellular%Chemokine,CCL28%Hypoxia inducible factor-1 α%Migration
目的 探讨趋化因子CCL28在缺氧诱导肝癌细胞侵袭中的作用. 方法 50份肝细胞肝癌标本来自肝细胞肝癌患者,肝癌细胞株HepG2、HCCLM3为实验室冻存.用实时定量聚合酶链反应检测肝癌组织中缺氧诱导因子-1 α(HIF-1 α)和CCL28的mRNA水平.实时定量聚合酶链反应、Western blot和ELISA检测不同缺氧时间处理后肝癌细胞HepG2、HCCLM3中HIF-1α和CCL28的mRNA和蛋白表达水平.构建CCL28-siRNA下调HCCLM3细胞CCL28表达后,Transwell观察缺氧诱导的肝癌HCCLM3细胞侵袭改变.用x2检验分析HIF-1α和CCL28表达水平与临床资料相关性,Spearman双变量相关性分析HIF-1α和CCL28表达水平相关性,用单因素方差分析组间比较. 结果 HIF-1α mRNA在肝癌组织中相对表达量为0.065土0.098,CCL28 mRNA水平为0.025±0.075,Spearman双变量分析显示肝癌组织中HIF-1 α与CCL28表达水平呈高度相关性(r=0.595,P＜0.01),二者表达水平增高皆与术后复发相关(P=0.011,P=0.019).缺氧培养肝癌HepG2和HCCLM3细胞CCL28表达增高并呈时间依赖性,HepG2:HIF-1αF=873.5,CCL28 F=151.6;HCCLM3..HIF-1αF=964.5,CCL28F=285.8,P值均＜0.01.SiRNA抑制CCL28表达后,缺氧条件下HCCLM3细胞侵袭数目为(43.2±5.4)个/室、空白对照组为(54.6±9.5)个/室、阴性对照组为(58.0土3.9)个/室,缺氧条件下HCCLM3细胞侵袭细胞数比空白对照组和阴性对照组减少,P=0.011.结论 趋化因子CCL28在缺氧条件下高表达并参与缺氧诱导肝癌细胞侵袭过程.
目的 探討趨化因子CCL28在缺氧誘導肝癌細胞侵襲中的作用. 方法 50份肝細胞肝癌標本來自肝細胞肝癌患者,肝癌細胞株HepG2、HCCLM3為實驗室凍存.用實時定量聚閤酶鏈反應檢測肝癌組織中缺氧誘導因子-1 α(HIF-1 α)和CCL28的mRNA水平.實時定量聚閤酶鏈反應、Western blot和ELISA檢測不同缺氧時間處理後肝癌細胞HepG2、HCCLM3中HIF-1α和CCL28的mRNA和蛋白錶達水平.構建CCL28-siRNA下調HCCLM3細胞CCL28錶達後,Transwell觀察缺氧誘導的肝癌HCCLM3細胞侵襲改變.用x2檢驗分析HIF-1α和CCL28錶達水平與臨床資料相關性,Spearman雙變量相關性分析HIF-1α和CCL28錶達水平相關性,用單因素方差分析組間比較. 結果 HIF-1α mRNA在肝癌組織中相對錶達量為0.065土0.098,CCL28 mRNA水平為0.025±0.075,Spearman雙變量分析顯示肝癌組織中HIF-1 α與CCL28錶達水平呈高度相關性(r=0.595,P＜0.01),二者錶達水平增高皆與術後複髮相關(P=0.011,P=0.019).缺氧培養肝癌HepG2和HCCLM3細胞CCL28錶達增高併呈時間依賴性,HepG2:HIF-1αF=873.5,CCL28 F=151.6;HCCLM3..HIF-1αF=964.5,CCL28F=285.8,P值均＜0.01.SiRNA抑製CCL28錶達後,缺氧條件下HCCLM3細胞侵襲數目為(43.2±5.4)箇/室、空白對照組為(54.6±9.5)箇/室、陰性對照組為(58.0土3.9)箇/室,缺氧條件下HCCLM3細胞侵襲細胞數比空白對照組和陰性對照組減少,P=0.011.結論 趨化因子CCL28在缺氧條件下高錶達併參與缺氧誘導肝癌細胞侵襲過程.
목적 탐토추화인자CCL28재결양유도간암세포침습중적작용. 방법 50빈간세포간암표본래자간세포간암환자,간암세포주HepG2、HCCLM3위실험실동존.용실시정량취합매련반응검측간암조직중결양유도인자-1 α(HIF-1 α)화CCL28적mRNA수평.실시정량취합매련반응、Western blot화ELISA검측불동결양시간처리후간암세포HepG2、HCCLM3중HIF-1α화CCL28적mRNA화단백표체수평.구건CCL28-siRNA하조HCCLM3세포CCL28표체후,Transwell관찰결양유도적간암HCCLM3세포침습개변.용x2검험분석HIF-1α화CCL28표체수평여림상자료상관성,Spearman쌍변량상관성분석HIF-1α화CCL28표체수평상관성,용단인소방차분석조간비교. 결과 HIF-1α mRNA재간암조직중상대표체량위0.065토0.098,CCL28 mRNA수평위0.025±0.075,Spearman쌍변량분석현시간암조직중HIF-1 α여CCL28표체수평정고도상관성(r=0.595,P＜0.01),이자표체수평증고개여술후복발상관(P=0.011,P=0.019).결양배양간암HepG2화HCCLM3세포CCL28표체증고병정시간의뢰성,HepG2:HIF-1αF=873.5,CCL28 F=151.6;HCCLM3..HIF-1αF=964.5,CCL28F=285.8,P치균＜0.01.SiRNA억제CCL28표체후,결양조건하HCCLM3세포침습수목위(43.2±5.4)개/실、공백대조조위(54.6±9.5)개/실、음성대조조위(58.0토3.9)개/실,결양조건하HCCLM3세포침습세포수비공백대조조화음성대조조감소,P=0.011.결론 추화인자CCL28재결양조건하고표체병삼여결양유도간암세포침습과정.
Objective To investigate the role of CCL28 in hypoxia-induced cell migration of hepatocellular carcinoma (HCC).Methods Resected liver tissues from 50 HCC patients were subjected to real-time (rt)-PCR analysis to evaluate the mRNA expression levels of the hypoxia-induced factor HIF-1α and the chemokine CCL28.Patient data on treatment and outcome were analyzed.The human HCC cell lines HepG2 and HCCLM3 were used to investigate effects of hypoxic conditions on HIF-1α and CCL28 expressions by rtPCR,western blotting,and enzyme-linked immunoassay.The CCL28-mediated effects of hypoxic conditions on cell mobility and invasion were assessed by trans-well and matrigel assays,respectively,in HCCLM3 with CCL28 expression silenced by small-interferring (si)RNA transfection.Spearman's rank test was used to assess the correlation between CCL28 and effects on disease-and treatment-related factors.Results The mRNA levels of CCL28 (0.025 ± 0.075) were found to be strongly correlated with HIF-1α(0.065 ± 0.098) in human clinical samples of HCC (r =0.595,P ＜ 0.01),with higher expressions of both related to recurrence after surgery (P=0.011 and 0.019,respectively).In vitro hypoxic conditions stimulated HIF-1α and CCL28 expression in a time-dependent manner in both HepG2 (HIF-1α:F=873.5; CCL28:F=151.6) and HCCLM3 (HIF-1α:F =964.5; CCL28:F =285.8) (allP ＜ 0.01).siRNA inhibition of CCL28 in HCCLM3 cells led to a significant reduction in hypoxia-induced invasion and migration (all P =0.011).Conclusion Chemokine CCL28 expression is up-regulated in human HCC and under in vitro hypoxic conditions,and may play an important role in hypoxia-induced HCC migration and invasion.
