中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2015年
1期
5-10
,共6页
畅秀丽%王黎明%臧梦雅%荣维淇%吴志远%刘立国%杜君%刘金宝%吴健雄
暢秀麗%王黎明%臧夢雅%榮維淇%吳誌遠%劉立國%杜君%劉金寶%吳健雄
창수려%왕려명%장몽아%영유기%오지원%류입국%두군%류금보%오건웅
肝肿瘤%肿瘤侵润%炎症趋化因子CCL20%受体,CCR6%Th17细胞
肝腫瘤%腫瘤侵潤%炎癥趨化因子CCL20%受體,CCR6%Th17細胞
간종류%종류침윤%염증추화인자CCL20%수체,CCR6%Th17세포
Liver Neoplasms%Neoplasm invasiveness%Chemokine CCL20%Receptors,CCR6%Th17 cells
目的 探讨CCL20-CCR6-Th17轴在肝细胞癌血管浸润转移中的作用.方法 采用SYBR Green实时定量PCR测定人肝细胞系L-02、肝细胞癌细胞系Hep3B、Huh7和HepG2中CCL20mRNA的表达,采用酶联免疫吸附试验(ELISA)检测上清中CCL20蛋白的分泌,Transwell迁移实验检测肝细胞癌细胞系对人外周血单个核细胞的趋化作用.采用ELISA定量检测93例肝细胞癌患者(转移组51例,无转移组42例)术前外周血中白细胞介素1α(IL-1α)、IL-1β 、IL-6 、IL-8 、IL-10、IL-17、IL-23、γ-干扰素、肿瘤坏死因子α和CCL20的水平,采用SYBR Green RT-PCR检测41例肝细胞癌癌组织及其相应的癌旁组织中CCL20和CCR6的表达水平,免疫组织化学染色检测肝细胞癌及其对应的癌旁组织和正常人供肝组织中CCL20的表达水平.结果 人肝细胞癌细胞系中CCL20的表达水平高于正常肝细胞,对外周血表达CCR6的T细胞具有趋化作用.93例肝细胞癌患者血清中CCL20蛋白的表达水平为(38.2±28.4) pg/ml,高于肝血管瘤患者[(7.8±17.8) pg/ml,P＜0.01].肝细胞癌患者血清中CCL20蛋白的表达水平与肿瘤直径呈正相关(r=0.32,P=0.0018).41例肝细胞癌组织CCL20 mRNA的表达水平高于癌旁组织(P＜0.05),CCL20蛋白主要在肝细胞癌细胞质中表达,某些浸润的免疫细胞也有一定程度表达.多因素分析结果显示,血清中IL-17和CCL20水平是影响肝细胞癌患者发生血管浸润转移的独立影响因素(均P ＜0.05).41例肝细胞癌患者癌组织及对应的癌旁组织中CCL20 mRNA的表达水平与肝细胞癌发生血管浸润转移均无关(均P＞0.05).在发生血管浸润转移患者的癌组织及其对应的癌旁组织中CCR6 mRNA的表达水平分别为5.75 (1.79,19.13)和7.99(4.49,19.54),均高于未发生血管浸润转移的患者[分别为1.69 (0.76,2.87)和3.58(1.84,4.32)],差异均有统计学意义(均P＜0.05).结论 CCL20/CCR6/Th 17轴可能促进肝细胞癌的血管浸润转移.
目的 探討CCL20-CCR6-Th17軸在肝細胞癌血管浸潤轉移中的作用.方法 採用SYBR Green實時定量PCR測定人肝細胞繫L-02、肝細胞癌細胞繫Hep3B、Huh7和HepG2中CCL20mRNA的錶達,採用酶聯免疫吸附試驗(ELISA)檢測上清中CCL20蛋白的分泌,Transwell遷移實驗檢測肝細胞癌細胞繫對人外週血單箇覈細胞的趨化作用.採用ELISA定量檢測93例肝細胞癌患者(轉移組51例,無轉移組42例)術前外週血中白細胞介素1α(IL-1α)、IL-1β 、IL-6 、IL-8 、IL-10、IL-17、IL-23、γ-榦擾素、腫瘤壞死因子α和CCL20的水平,採用SYBR Green RT-PCR檢測41例肝細胞癌癌組織及其相應的癌徬組織中CCL20和CCR6的錶達水平,免疫組織化學染色檢測肝細胞癌及其對應的癌徬組織和正常人供肝組織中CCL20的錶達水平.結果 人肝細胞癌細胞繫中CCL20的錶達水平高于正常肝細胞,對外週血錶達CCR6的T細胞具有趨化作用.93例肝細胞癌患者血清中CCL20蛋白的錶達水平為(38.2±28.4) pg/ml,高于肝血管瘤患者[(7.8±17.8) pg/ml,P＜0.01].肝細胞癌患者血清中CCL20蛋白的錶達水平與腫瘤直徑呈正相關(r=0.32,P=0.0018).41例肝細胞癌組織CCL20 mRNA的錶達水平高于癌徬組織(P＜0.05),CCL20蛋白主要在肝細胞癌細胞質中錶達,某些浸潤的免疫細胞也有一定程度錶達.多因素分析結果顯示,血清中IL-17和CCL20水平是影響肝細胞癌患者髮生血管浸潤轉移的獨立影響因素(均P ＜0.05).41例肝細胞癌患者癌組織及對應的癌徬組織中CCL20 mRNA的錶達水平與肝細胞癌髮生血管浸潤轉移均無關(均P＞0.05).在髮生血管浸潤轉移患者的癌組織及其對應的癌徬組織中CCR6 mRNA的錶達水平分彆為5.75 (1.79,19.13)和7.99(4.49,19.54),均高于未髮生血管浸潤轉移的患者[分彆為1.69 (0.76,2.87)和3.58(1.84,4.32)],差異均有統計學意義(均P＜0.05).結論 CCL20/CCR6/Th 17軸可能促進肝細胞癌的血管浸潤轉移.
목적 탐토CCL20-CCR6-Th17축재간세포암혈관침윤전이중적작용.방법 채용SYBR Green실시정량PCR측정인간세포계L-02、간세포암세포계Hep3B、Huh7화HepG2중CCL20mRNA적표체,채용매련면역흡부시험(ELISA)검측상청중CCL20단백적분비,Transwell천이실험검측간세포암세포계대인외주혈단개핵세포적추화작용.채용ELISA정량검측93례간세포암환자(전이조51례,무전이조42례)술전외주혈중백세포개소1α(IL-1α)、IL-1β 、IL-6 、IL-8 、IL-10、IL-17、IL-23、γ-간우소、종류배사인자α화CCL20적수평,채용SYBR Green RT-PCR검측41례간세포암암조직급기상응적암방조직중CCL20화CCR6적표체수평,면역조직화학염색검측간세포암급기대응적암방조직화정상인공간조직중CCL20적표체수평.결과 인간세포암세포계중CCL20적표체수평고우정상간세포,대외주혈표체CCR6적T세포구유추화작용.93례간세포암환자혈청중CCL20단백적표체수평위(38.2±28.4) pg/ml,고우간혈관류환자[(7.8±17.8) pg/ml,P＜0.01].간세포암환자혈청중CCL20단백적표체수평여종류직경정정상관(r=0.32,P=0.0018).41례간세포암조직CCL20 mRNA적표체수평고우암방조직(P＜0.05),CCL20단백주요재간세포암세포질중표체,모사침윤적면역세포야유일정정도표체.다인소분석결과현시,혈청중IL-17화CCL20수평시영향간세포암환자발생혈관침윤전이적독립영향인소(균P ＜0.05).41례간세포암환자암조직급대응적암방조직중CCL20 mRNA적표체수평여간세포암발생혈관침윤전이균무관(균P＞0.05).재발생혈관침윤전이환자적암조직급기대응적암방조직중CCR6 mRNA적표체수평분별위5.75 (1.79,19.13)화7.99(4.49,19.54),균고우미발생혈관침윤전이적환자[분별위1.69 (0.76,2.87)화3.58(1.84,4.32)],차이균유통계학의의(균P＜0.05).결론 CCL20/CCR6/Th 17축가능촉진간세포암적혈관침윤전이.
Objective To investigate the role of CCL20/CCR6/Th17 axis in vascular invasion and metastasis of primary hepatocellular carcinoma (HCC).Methods Expression levels of CCL20 mRNA in the normal human liver cell line L-02,and human hepatocellular carcinoma cell lines Hep3B,Huh7 and HepG2 were quantified by using SYBR green real time PCR.CCL20 secretions from these cell lines were quantified by using ELISA.The chemotactic effect of HCC cell line Hep3B on human peripheral blood mononuclear cells was determined by using transwell chemotaxis assay.Pre-therapy serum levels of IL-1 α,IL-1β,IL-6,IL-8,IL-10,IL-17,IL-23,IFN-γ,TNF-α and CCL20 in 93 patients with HCC were measured by using 9-plex array and ELISA.All the patients were chronic hepatitis B virus associated HCC,and 51 cases were those with vascular invasion and metastasis (metastasis group) and 42 cases were not (non-metastasis group).CCL20 and CCR6 mRNA expressions in the HCC and tumor-adjacent tissues were determined by using SYBR Green real time PCR in 41 patients,among them,20 cases were from the group of patients with metastasis and 21 cases were from the group of patients without metastasis.The CCL20 expression was further determined by immunohistochemistry.Results The HCC cell lines expressed and secreted higher amount of CCL20,which effectively recruited CCR6 + T cells.Pre-therapy serum levels of CCL20 in 93 HCC patients were (38.2 ± 28.4) pg/ml,significantly increased than those with benign hepatic hemangiomas [(7.8 ± 17.8)pg/ml,P ＜0.01].In addition,the serum levels of CCL20 were positively correlated with the tumor diameters in HCC patients (r =0.32,P =0.0018).CCL20 was dominantly expressed in the cytoplasm in HCC cells,and it was also expressed by some infiltrating immune cells.The mRNA expression levels of CCL20 of the tumor tissues were significantly higher than that in the tumor-adjacent tissues (P ＜ 0.05).Multivariate logistic regression analysis showed that serum levels of IL-17 and CCL20 were independent risk factors of metastasis in HCC patients (P ＜ 0.05 for both).CCL20 mRNA showed no statistically significant differences between patients with metastasis and without metastasis in both tumor tissues and tumor-adjacent tissues (P ＞ 0.05 for both).But the patients with metastasis showed significantly higher expressions of CCR6 both in their tumor [5.75 (1.79,19.13)] and tumor-adjacent tissues [7.99 (4.49,19.54)] than those with non-metastasis [1.69 (0.76,2.87) and 3.58 (1.84,4.32),P ＜ 0.05 for both].Conclusion CCL20/CCR6/Th17 axis may promote vascular invasion and metastasis hepatocellular carcinoma.
