目的探讨CCL18-PITPNM3(CC chemokine ligand 18-phosphatidylinositol transfer protein 3,CCL18-PITPNM3)配体受体轴在头颈鳞状细胞癌侵袭转移中的作用及其分子机制。方法采用人重组蛋白CCL18处理头颈鳞状细胞癌Tu686、6-10B细胞,siRN...目的探讨CCL18-PITPNM3(CC chemokine ligand 18-phosphatidylinositol transfer protein 3,CCL18-PITPNM3)配体受体轴在头颈鳞状细胞癌侵袭转移中的作用及其分子机制。方法采用人重组蛋白CCL18处理头颈鳞状细胞癌Tu686、6-10B细胞,siRNA下调PITPNM3的表达,通过CCK-8(cell counting kit-8)、平板克隆实验、流式周期检测生长增殖能力的变化,划痕愈合实验、Transwell侵袭小室实验检测体外迁移侵袭能力的改变,qRT-PCR、Western blot检测EMT分子标志物的表达情况。结果①rhCCL18处理头颈鳞状细胞癌Tu686、6-10B细胞后,细胞划痕愈合率增加,穿过Transwell聚碳酸酯膜的细胞明显增多,rhCCL18处理siRNA下调PITPNM3的Tu686、6-10B细胞,下调组细胞的迁移和侵袭能力较亲本细胞处理组明显减弱;②rhCCL18处理头颈鳞状细胞癌Tu686、6-10B细胞,mRNA水平E-cadherin表达降低,Vimentin、N-cadherin、Fibronectin表达升高;蛋白质水平E-cadherin表达降低,Vimentin表达升高。下调两株细胞的PITPNM3表达后rhCCL18再次处理,E-cadherin下调和Vimentin、N-cadherin、Fibronectin上调均未显示出亲本细胞的明显趋势;③rhCCL18处理和下调PITPNM3对Tu686、6-10B 6组细胞的生存率、增殖及细胞周期无明显变化。结论CCL18-PITPNM3配体受体轴可促进头颈鳞状细胞癌的体外侵袭转移能力,可能与EMT转化相关。展开更多
A previous study indicated that C–C chemokine(C–C motif)ligand 18(CCL18)is capable of inducing tumor cell invasion and metastasis by interacting with receptor membrane-associated phosphatidylinositol transfer protei...A previous study indicated that C–C chemokine(C–C motif)ligand 18(CCL18)is capable of inducing tumor cell invasion and metastasis by interacting with receptor membrane-associated phosphatidylinositol transfer protein 3(PITPNM3)in breast cancer cells.The present study aims to investigate the correlation between the PITPNM3 expression and metastasis in hepatocellular carcinoma(HCC).Real-time quantitative polymerase chain reaction and Western blot were performed to detect the expression pattern of PITPNM3 in patient samples and HCC cell lines.Wound-healing and transwell chamber assays were performed to assess the migration and invasiveness of HCC cells,and the activation of the signaling protein downstream of PITPNM3 was also detected by Western blot and immunofluorescence.The results revealed that PITPNM3 was upregulated in HCC tissue compared to matched normal liver tissue.Silencing the expression of PITPNM3 by specific siRNAs markedly attenuated the invasive and metastatic abilities of HCC cells,whereas the upregulation of PITPNM3 significantly increased HCC cell mobility.Furthermore,inhibiting the expression of PITPNM3 suppressed the activation of Pyk2,FAK,and Src,while overexpression of PITPNM3enhanced the phosphorylation of FAK and Src in HCC cells.Besides,suppression of Pyk2 can also impair the clustering of integrin.These results imply that PITPNM3 is a vital determinant of HCC migration and invasion.展开更多
基金supported by the National Key Basic Research Program of China(2010CB912800,2011CB504203)the National Natural Science Foundation of China(81102022,81230060,81261140373,81000917,and 81372819)+6 种基金the Foundationfor the Young Teachers in the Higher Education Institutions of China(20110171120082)the National S&T Major Special Project on New Drug Innovation of China(2011ZX09102-010-02)the Science Foundation of Guangdong Province(S2012030006287)the Translational Medicine Public Platform of Guangdong Province(4202037)the Foundation of the Ministry of Education of China(20120171110075)funding from Sun Yat-Sen University(13ykzd14)the grant[2013]163 from Key Laboratory of Malignant Tumor Molecular Mechanism and Translational Medicineof Guangzhou Bureau of Science and Information Technology
文摘A previous study indicated that C–C chemokine(C–C motif)ligand 18(CCL18)is capable of inducing tumor cell invasion and metastasis by interacting with receptor membrane-associated phosphatidylinositol transfer protein 3(PITPNM3)in breast cancer cells.The present study aims to investigate the correlation between the PITPNM3 expression and metastasis in hepatocellular carcinoma(HCC).Real-time quantitative polymerase chain reaction and Western blot were performed to detect the expression pattern of PITPNM3 in patient samples and HCC cell lines.Wound-healing and transwell chamber assays were performed to assess the migration and invasiveness of HCC cells,and the activation of the signaling protein downstream of PITPNM3 was also detected by Western blot and immunofluorescence.The results revealed that PITPNM3 was upregulated in HCC tissue compared to matched normal liver tissue.Silencing the expression of PITPNM3 by specific siRNAs markedly attenuated the invasive and metastatic abilities of HCC cells,whereas the upregulation of PITPNM3 significantly increased HCC cell mobility.Furthermore,inhibiting the expression of PITPNM3 suppressed the activation of Pyk2,FAK,and Src,while overexpression of PITPNM3enhanced the phosphorylation of FAK and Src in HCC cells.Besides,suppression of Pyk2 can also impair the clustering of integrin.These results imply that PITPNM3 is a vital determinant of HCC migration and invasion.
