趋化因子2对乳腺癌细胞MCF-7趋化活性及趋化因子5mRNA表达的影响

Objective To observe the effects of chemokine 2 (CCL2) on the expression of chemokine 5 ( CCL5) mRNA and chemotactic activity of MCF-7 cells. Methods MCF-7 cells were treated with different concentrations of CCL2, the expression of CCL5 mRNA was detected by using real-time fluorescence quantitative polymerase chain reaction ( RTFQ-PCR), and the chemotactic activity of MCF-7 was measured by using chemotaxis chamber method. Results When the concentration of exogenous CCL2 was 200 μg/L, the MCF-7 cells expressed the highest CCL5 mRNA (15. 22 ± 2. 3, P ＜0. 01). With the prolongation of CCL2 action time, the expression levels of CCL5 mRNA were increased. There was a positive correlation between the chemotactic activity of MCF-7 cells and the concentration of CCL2. When the concentration of exogenous CCL2 was 300 μg/L, the number of penetrating cells was the greatest (88.00 ±11. 53, P ＜0. 01). With the prolongation of CCL2 action time, the chemotactic activity of MCF-7 cells was enhanced. When the action time was 30 h, the number of penetrating cells was the greatest (81.00 ±9. 54, P ＜ 0.05 ). Conclusion Exogenous CCL2 could increase the expression of CCL5 mRNA and the chemotactic activity of MCF-7 cells.     

趋化因子2对乳腺癌细胞MCF-7趋化活性及趋化因子5mRNA表达的影响

Objective To observe the effects of chemokine 2 (CCL2) on the expression of chemokine 5 ( CCL5) mRNA and chemotactic activity of MCF-7 cells. Methods MCF-7 cells were treated with different concentrations of CCL2, the expression of CCL5 mRNA was detected by using real-time fluorescence quantitative polymerase chain reaction ( RTFQ-PCR), and the chemotactic activity of MCF-7 was measured by using chemotaxis chamber method. Results When the concentration of exogenous CCL2 was 200 μg/L, the MCF-7 cells expressed the highest CCL5 mRNA (15. 22 ± 2. 3, P ＜0. 01). With the prolongation of CCL2 action time, the expression levels of CCL5 mRNA were increased. There was a positive correlation between the chemotactic activity of MCF-7 cells and the concentration of CCL2. When the concentration of exogenous CCL2 was 300 μg/L, the number of penetrating cells was the greatest (88.00 ±11. 53, P ＜0. 01). With the prolongation of CCL2 action time, the chemotactic activity of MCF-7 cells was enhanced. When the action time was 30 h, the number of penetrating cells was the greatest (81.00 ±9. 54, P ＜ 0.05 ). Conclusion Exogenous CCL2 could increase the expression of CCL5 mRNA and the chemotactic activity of MCF-7 cells.     

慢病毒介导CCL5-siRNA对乳腺癌细胞生物学行为的影响

Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.     

慢病毒介导CCL5-siRNA对乳腺癌细胞生物学行为的影响

Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.     

慢病毒介导CCL5-siRNA对乳腺癌细胞生物学行为的影响

Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.     

慢病毒介导CCL5-siRNA对乳腺癌细胞生物学行为的影响

Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.     

慢病毒介导CCL5-siRNA对乳腺癌细胞生物学行为的影响

Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.     

CCL5对MCF-7中细胞增殖能力及CD44~+/CD24~-亚群比例的影响

目的 观察外源性趋化因子5(CCL5)对人乳腺癌MCF-7细胞增殖及CD44+/CD24-亚群比例的影响,探讨其在乳腺癌进展过程中的作用.方法 噻唑蓝(MTT)比色法检测浓度为10、50、100、500 μg/L的外源性rhCCL5(recombinant human CCL5)作用0、24、48 h后MCF-7细胞增殖水平的变化,流式细胞仪检测CD44~+/CD24~-亚群比例的变化.结果 rhCCL5作用48 h后MCF-7细胞增殖水平高于对照组(P＜0.05),并呈浓度依赖趋势;且高于作用24、0 h后细胞的增殖水平(P＜0.05).50、500μg/L rhCCL5处理MCF-7细胞后升高了细胞株中CD44~+/CD24~-亚群的比例,分别为对照组的5.68和6.05倍(P＜0.05).结论 外源性CCL5可以明显升高MCF-7细胞的增殖水平以及CD44~+/CD24~-亚群的比例,这种作用可能是其诱导乳腺癌进展的机制之一.     

siRNA干扰Ezrin表达对CCL5介导人乳腺癌细胞侵袭能力的影响

目的 探讨Ezrin在趋化因子CCL5促进人乳腺癌细胞MCF-7转移中的作用及其机制.方法 采用小干扰RNA(siRNA)方法下调人乳腺癌细胞中Ezrin的表达,Western blot检测siRNA对Ezrin mRNA和蛋白表达水平下调的变化.干扰后,趋化小室检测MCF-7细胞在不同浓度CCL5作用下的侵袭活性;Western blot检测细胞中总的Ezrin蛋白及磷酸化的Ezrin蛋白表达随CCL5趋化时间的变化.结果 与MCF-7细胞和转染空白质粒pSUPER细胞比较,在CCL5趋化作用下MCF-7细胞Ezrin蛋白的表达无明显增加(P>0.05),干扰后,在50μg/L CCL5作用下,MCF-7细胞表现出侵袭活性最强(F=31.62,P<0.01),但仍较对照组明显降低(F=41.96,P<0.01).结论 趋化因子CCL5促进人乳腺癌细胞MCF-7的趋化与侵袭,Ezrin蛋白的激活在此过程中起着一定的作用.     

siRNA干扰Ezrin表达对CCL5介导人乳腺癌细胞侵袭能力的影响

目的 探讨Ezrin在趋化因子CCL5促进人乳腺癌细胞MCF-7转移中的作用及其机制.方法 采用小干扰RNA(siRNA)方法下调人乳腺癌细胞中Ezrin的表达,Western blot检测siRNA对Ezrin mRNA和蛋白表达水平下调的变化.干扰后,趋化小室检测MCF-7细胞在不同浓度CCL5作用下的侵袭活性;Western blot检测细胞中总的Ezrin蛋白及磷酸化的Ezrin蛋白表达随CCL5趋化时间的变化.结果 与MCF-7细胞和转染空白质粒pSUPER细胞比较,在CCL5趋化作用下MCF-7细胞Ezrin蛋白的表达无明显增加(P>0.05),干扰后,在50μg/L CCL5作用下,MCF-7细胞表现出侵袭活性最强(F=31.62,P<0.01),但仍较对照组明显降低(F=41.96,P<0.01).结论 趋化因子CCL5促进人乳腺癌细胞MCF-7的趋化与侵袭,Ezrin蛋白的激活在此过程中起着一定的作用.     

siRNA干扰Ezrin表达对CCL5介导人乳腺癌细胞侵袭能力的影响

目的 探讨Ezrin在趋化因子CCL5促进人乳腺癌细胞MCF-7转移中的作用及其机制.方法 采用小干扰RNA(siRNA)方法下调人乳腺癌细胞中Ezrin的表达,Western blot检测siRNA对Ezrin mRNA和蛋白表达水平下调的变化.干扰后,趋化小室检测MCF-7细胞在不同浓度CCL5作用下的侵袭活性;Western blot检测细胞中总的Ezrin蛋白及磷酸化的Ezrin蛋白表达随CCL5趋化时间的变化.结果 与MCF-7细胞和转染空白质粒pSUPER细胞比较,在CCL5趋化作用下MCF-7细胞Ezrin蛋白的表达无明显增加(P>0.05),干扰后,在50μg/L CCL5作用下,MCF-7细胞表现出侵袭活性最强(F=31.62,P<0.01),但仍较对照组明显降低(F=41.96,P<0.01).结论 趋化因子CCL5促进人乳腺癌细胞MCF-7的趋化与侵袭,Ezrin蛋白的激活在此过程中起着一定的作用.     

次黄嘌呤单核苷酸脱氢酶抑制剂对人外周髓样树突状细胞趋化、迁移和吞噬功能的影响研究

目的 探讨次黄嘌呤单核苷酸脱氢酶抑制剂(Inosine monophosphate dehydrogenase inhibitor,IMPDHI)对人外周髓样树突状细胞(Myeloid dendritic cells,MDC)趋化、迁移、吞噬功能的影响.方法 新鲜外周血单个核细胞来源于健康志愿者(N=15).实验组加入不同浓度IMPDHI,流式细胞仪分析MDC表面趋化因子受体表达水平.于transwell小室实验中,加入不同的化学因子,经Lin-1/CD11c/HLA-DR染色后,流式细胞仪计数,以迁移细胞的百分比表示迁移能力.分离树突状细胞抗原-1+(Blood dendritic cell antigen-1,BDCA-1+)细胞后,以甘露糖受体作为介导,流式细胞仪测定BDCA-1+细胞中FITC标记的右旋糖酐的荧光值表示吞噬能力.结果 (1)趋化、迁移功能:与对照组相比,实验组MDC的趋化因子受体CCR1表达水平明显升高(17.02±3.23～30.63±9.13,P＜0.05);CCR3表达水平(10.26±2.25～5.81±0.97,P＜0.05)和CCR7表达水平(9.56±1.84～5.18±0.60,P＜0.05)明显下降.实验组MDC对炎性化学因子CCL2、CCL3、CCL4、CCL7、CXCL12的趋化能力明显增强(P＜0.05),对淋巴器官性化学因子CCL19、CCL20、CCL21、CXCL11的趋化能力无明显改变(P＞0.05).(2)吞噬功能:实验组MDC的吞噬能力明显强于对照组(P＜0.05).结论 IMPDHI增强MDC吞噬抗原的能力,通过提高MDC炎性化学因子受体的表达水平和增强其对炎性化学因子的趋化能力,抑制MDC对淋巴器官的趋化、迁移能力.     

肿瘤相关钙信号传导蛋白-2基因对人乳腺癌细胞黏附和侵袭力的影响

目的 观察肿瘤相关钙信号传导蛋白-2(TROP-2)基因小干扰RNA (siRNA)对乳腺癌细胞黏附和侵袭力的影响.方法 培养人乳腺癌Bcap-37、LCC1、MCF-7、MDA-MB-231、MDA-MB-435、MDA-MB-468及ZR75-1细胞株,以荧光实时定量聚合酶链反应(PCR)方法检测TROP-2基因mRNA表达;筛选出TROP-2表达最高者.采用TROP4基因siRNA转染乳腺癌细胞,分别以荧光实时定量RT-PCR和免疫荧光方法观察TROP-2基因mRNA和蛋白水平,然后以噻唑蓝(MTT)比色法检测细胞黏附性,以Transwell方法检测癌细胞侵袭能力.结果 人乳腺癌Bcap-37、LCC1、MCF-7、MDA-MB-231、MDA-MB-35、MDA-MB-468及ZR75-1细胞株TROP-2 mRNA分别是1.362±0.057、2.207±0.056、2.997±0.052、0.136±0.045、0.122±0.025、0.194±0.028和2.706±0.039,以MCF-7细胞最高;以TROP-2 siRNA转染乳腺癌MCF-7细胞后,癌细胞TROP-2基因mRNA和蛋白水平明显下降,且呈浓度依赖性;黏附实验结果显示,5、10、20 nmol/L siRNA组黏附率分别为(52.9±2.5)％、(25.6±2.3)％、(12.8±2.2)％(P＜0.01);Transwell实验结果显示,5、10和20 mol/L siRNA组穿过滤膜的细胞分别为78±17、39±15、19±16,而对照组分别为136±25、139±21(P＜0.01).结论 TROP-2基因在乳腺癌细胞黏附和侵袭中发挥着重要作用;以siRNA转染乳腺癌细胞,可抑制乳腺癌细胞黏附和侵袭能力.     

慢病毒介导CCL5-siRNA对乳腺癌细胞生物学行为的影响

目的 探讨CCL5-siRNA对高转移性人乳腺癌细胞生物学行为的影响.方法 合成特异性CCL5-siRNA并克隆入慢病毒表达载体pGCSIL-GFP,将重组CCL5-siRNA慢病毒感染高转移性人乳腺癌细胞系MDA-MB-231设为KD组,另设阴性对照组和未感染组;分别采用实时定量聚合酶链反应和Western blot方法检测CCL5-siRNA对CCL5表达的作用;用噻唑蓝比色法、流式细胞术、克隆形成试验和Boyden小窒穿透试验观察细胞的增殖、周期、体外聚集和侵袭力的改变.结果 CCL5-siRNA慢病毒可显著降低MDA-MB-231细胞CCL5的表达,改变细胞形成克隆和运动侵袭的能力,但对细胞增殖的影响不明显.与阴性对照组(1.00±0.07)和未感染组(0.88±0.15)比较,KD组细胞CCL5 mRNA的表达下降为(0.18±0.03),P<0.01.在上样量相同的条件下,与阴性对照组(1.82±0.18)比较,KD组CCL5蛋白表达降低为(0.33±0.13),P<0.01.细胞克隆计数:与阴性对照组(0.97±0.09)和未感染组(1.04±0.07)比较,KD组细胞的克隆数降为(0.33±0.10),P<0.05;穿膜细胞计数:KD组(38±15)明显少于阴性对照组(77±11)和末感染组(69±9),P<0.05.嚷唑蓝比色实验和流式细胞分析提示:KD组细胞的增殖情况与阴性对照组、未感染组相比无明显差别.三组PI值分别为0.48±0.02、0.44±0.05及0.47±0.02(P>0.05).结论 慢病毒介导的CCL5-siRNA可显著抑制高转移性人乳腺癌细胞的克隆形成和运动侵袭能力,但对其增殖活性无明显影响.     

趋化因子受体5促进乳腺癌干细胞的趋化与侵袭

目的 观察趋化因子受体5(CCR5)对乳腺癌干细胞(CD44+CD24-/low)趋化和侵袭作用.方法 应用流式细胞仪分选获得乳腺癌干细胞,逆转录-聚合酶链反应(RT-PCR)和酶联免疫吸附试验(ELISA)检测乳腺癌MCF-72个SP侧群细胞(side population)6例样本CCR5 mRNA和蛋白质,在趋化因子5(CCL5)作用下通过趋化小室法检测2个SP侧群细胞趋化活性和侵袭活性,并与CCR5的封闭进行对照研究.结果 CCL5对MCF-7肿瘤干细胞有明显趋化[乳腺癌干细胞(86.0±14.8)个/HP与CD44+CD24+(72.0±13.5)个/HP,t=7.461,P<0.05]和侵袭作用[乳腺癌干细胞(25.0±8.3)个/HP与CD44+CD24+(16.0±5.4)个/HP,t=6.665,P<0.05].结论 CCB5表达能够促进乳腺癌干细胞趋化和侵袭.     

胃癌患者血清中CCL20和CCR6的表达及临床意义

目的 测定趋化因子CCL20及其受体CCR6在胃癌患者的血清中表达,并探讨CCL20和CCR6表达水平与胃癌的发生发展关系.方法 应用荧光定量PCR技术、流式细胞术和酶联免疫吸附法测定50例胃癌患者和30例健康对照者外周血中CCL20和CCR6的mRNA和蛋白表达水平.结果 胃癌患者CCL20和CCR6的mRNA表达均显著高于健康对照组(P<0.01);胃癌患者外周血中CCL20和CCR6的蛋白表达分别为(45.4±10.9)pg/mL和(7.11±1.03)%,显著高于健康人的(18.6±4.7)pg/mL和(1.83±0.43)%(P<0.01),且其升高程度与胃癌临床分期有明显关系.结论 胃癌患者CCL20和CCR6的表达与胃癌的发生发展存在一定的相关性,可能参与胃癌的发病过程,推测其可能作为胃癌诊断、复发和转移的分子标志物.