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

目的 观察趋化因子2(CCL2)对MCF-7表达趋化因子5(CCL5)mRNA的影响,并观察CCL2对MCF-7趋化活性的影响.方法 使用不同浓度的CCL2作用于MCF-7细胞,通过实时荧光定量聚合酶链反应(RTFQ-PCR)测定不同时间CCL5 mRNA的表达,并使用趋化小室法检测CCL2作用下MCF-7趋化活性的改变.结果 当外源性CCL2浓度为200μg/L时,MCF-7中CCL5 mRNA相对表达量最大(15.22±2.32)(P＜0.01),随着作用时间的延长,CCL5 mRNA表达量增加;MCF-7的趋化活性与CCL2浓度呈正相关,当CCL2浓度为300μg/L时,穿膜细胞数最多(88.00±11.53)(P＜0.01);MCF-7的趋化活性与CCL2作用时间呈正相关,当CCL2作用时间为30 h时,穿膜细胞数最多(81.00±9.54)(P＜0.05).结论 加入外源性CCL2,MCF-7中CCL2 mRNA表达量增加,MCF-7趋化活性增强.

Abstract：

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.     

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.     

Caveolin-1 as Tumor Suppressor Gene in Breast Cancer

Purpose. Many articles have reported the caveolin-1 gene to be downregulated thus suggesting that it might be a candidate tumor suppressor gene in many tumors, but in bladder tumors the caveolin-1 gene was expressed and related to the pathological grade. We investigated the function of caveolin-1 protein in breast tumors. Methods. We introduced the caveolin-1 gene into a MCF-7 human breast cancer cell line, and examined its cell growth, cell viability, and anchorage-independent growth. Results. The caveolin-1 transfectants showed less proliferation than the vector control transfectants on the fourth day regarding the cell growth rate. In addition, the cell viability of the caveolin-1 transfectants was about 55% of the vector control. A soft agar assay of the caveolin-1 transfectants showed less growth with a lower number of colonies. Conclusion. In MCF-7 cells, the caveolin-1 gene may influence the tumor suppressor efficacy. Received: May 20, 2002 / Accepted: November 19, 2002 RID="*" ID="*" Reprint requests to: M. Hino, Niigata Cancer Center Hospital, 2-15-3 Kawagishi-cho, Niigata 951-8566, Japan     

灵菌红素通过促进Bim调控人乳腺癌细胞凋亡

目的探讨Bim在灵菌红素调控人乳腺癌MCF-7细胞增殖和凋亡中发挥的作用。方法培养人乳腺癌MCF-7细胞、采用灵菌红素处理MCF-7细胞,应用MTT检测细胞活力增殖及流式细胞计数检测细胞凋亡率,RT-PCR和Western blot检测Bim mRNA和蛋白表达情况。结果灵菌红素能抑制MCF-7细胞增殖,并诱导MCF-7细胞凋亡,效果呈时间和浓度依赖性(P0.05)。灵菌红素处理后MCF-7细胞S期显著减少、增殖指数下降,G1期显著增加(P0.05)。灵菌红素促进MCF-7细胞内促凋亡蛋白Bim表达升高,细胞凋亡率亦显著上升;沉默Bim能拮抗灵菌红素对MCF-7细胞凋亡的促进作用(P0.05)。结论灵菌红素通过上调Bim从而抑制人乳腺癌MCF-7细胞增殖和促进其凋亡。     

纳洛酮预处理对吗啡抑制人乳腺癌MCF-7细胞生长增殖的影响

目的观察纳洛酮预处理对吗啡抑制人乳腺癌MCF-7细胞生长增殖的影响。方法人乳腺癌MCF-7细胞培养至对数生长期,采用随机数字表法,将其分为四组:空白对照组(C组),10μmol/L吗啡组(M组),10μmol/L纳洛酮组(N组)及10μmol/L吗啡+10μmol/L纳洛酮组(MN组)。M组在培养液中加入吗啡,使吗啡的终浓度为10μmol/L;N组在培养液中加入纳洛酮,使其终浓度为10μmol/L;MN组则先在培养液中加入纳洛酮,使其浓度为10μmol/L,孵育30min后再将吗啡加入培养液中,使其终浓度为10μmol/L;对照组不加入任何药物。采用四唑盐(MTT)法和克隆形成实验观察细胞生长增殖的变化,流式细胞仪检测细胞周期分布和细胞凋亡率。结果C组与N组细胞活力、克隆形成率、细胞周期分布及细胞凋亡率差异无统计学意义;M组及MN组细胞的生长速度明显慢于C组,克隆形成率、细胞停滞在S期的比例明显低于C组,而细胞凋亡率、细胞停滞在G2/M期的比例明显高于C组(P0.05);M组与MN组细胞活力、克隆形成率、细胞周期分布及细胞凋亡率差异无统计学意义。结论在吗啡抑制人乳腺癌MCF-7细胞生长增殖、促进细胞凋亡的过程中,纳洛酮没有发挥拮抗作用,这说明吗啡抑制乳腺癌MCF-7细胞生长增殖的作用与阿片受体途径无关。     

MCF-7细胞系乳腺癌裸鼠模型病理学特点及生物学性状的研究

目的 探讨荷人乳腺癌裸鼠移植模型的病理学特点及生物学性状,为乳腺癌病因学、发病学研究提供可靠的工具.方法 在裸鼠右侧腋窝注射人乳腺癌MCF-7细胞悬液0.1 mL(5×10~8个/mL),建立乳腺癌裸鼠动物模型.混合干预组小鼠,肿瘤接种部位旁皮下注射瘦素0.2 mL(15×10~4 ng/mL)和雌激素0.2 mL(0.15 mg/mL),每天1次;雌激素组注射雌激素0.2 mL(O.15 mg/mL);瘦素组注射瘦素0.2 mL(15×10~4ng/mL);空白组注射0.9%氯化钠溶液,每日观察肿块生长情况及肿瘤体积变化.宰杀取材,观察肿瘤组织学特征.结果 (1)空白组肿瘤移植成功率为33.3%(10/30),瘦素组为46.7%(14/30),模型失败.混合干预组肿瘤移植成功率为96.7%(29/30),雌激素组为93.3%(28/30),两组间无明显差异(P>0.05);(2)混合干预组裸鼠平均肿瘤最长径明显优于雌激素组,差异具有统计学意义(P相似文献   

维生素E琥珀酸酯对乳腺癌荷瘤裸鼠的抑瘤作用

目的 检测维生素E琥珀酸酯（VES）对裸鼠荷乳腺癌MCF-7细胞的抑制作用。方法 裸鼠皮下接种MCF-7乳腺癌细胞建立荷瘤裸鼠模型。以150mg／kg（体重）剂量的VES治疗模型鼠共5周。处死动物后测量肿瘤大小。以流式细胞仪检测细胞周期变化和细胞表面Fas和FasL的表达；以免疫组化法检测肿瘤组织Fas和FasL的表达，并以TUNEL法检测细胞凋亡指数。结果 VES对荷乳腺癌裸鼠具有显著的增殖抑制作用。细胞周期分析显示，VES冶疗后肿瘤细胞表现为G0／G1期阻滞，肿瘤组织和细胞表面的Fas和FasL均出现上调，同时细胞的凋亡指数升高。结论 VES对于MCF-7乳腺癌荷瘤裸鼠具有强效的抑制作用，其机制与上调肿瘤细胞Fas／FasL表达，促进细胞凋亡有关。     

Effects of tamoxifen on growth and apoptosis of estrogen-dependent and -independent human breast cancer cells

Background: Apoptosis (“programmed cell death”) is an active process characterized by prominent nuclear changes and DNA cleavage, which distinguishes it from cellular necrosis. In this study we investigated whether tamoxifen (TAM) treatment of estrogen receptor ER(+) MCF-7 and ER(-) MDA-231 human breast cancer cells resulted in cytotoxicity and cellular changes typical of apoptosis. Methods: Cytotoxicity was measured using a tetrazolium assay. Cellular morphologic changes were observed using transmission electron microscopy. DNA cleavage was assessed using 1.6% agarose gel electrophoresis and was also quantitated biochemically. Results: Exposure of cells to TAM for 24 h resulted in dose- dependent cytotoxicity, and MCF-7 cells were somewhat more sensitive to TAM. TAM induced chromatin condensation around the nuclear periphery in both cell lines, changes typical of apoptosis. TAM-induced cytotoxicity correlated with dose-dependent DNA cleavage, which showed the characteristic “internucleosomal ladder.” DNA cleavage occurred at a slightly lower TAM dose and occurred somewhat sooner in MCF-7 cells. TAM-induced DNA cleavage in MCF-7 cells was inhibited by the protein synthesis inhibitor cycloheximide, the RNA synthesis inhibitor actinomycin D, and by 17β-estradiol. However, in MDA-231 cells, DNA cleavage was inhibited by cycloheximide, partially but not significantly inhibited by actinomycin D, and not inhibited by 17β-estradiol. Conclusions: TAM induces typical apoptosis in ER(+) or ER (-) human breast cancer cells. TAM induction of apoptosis in MCF-7 cells involves the estrogen receptor, and requires the synthesis of new protein and mRNA. TAM induction of apoptosis in MDA-231 cells depends primarily on protein synthesis. TAM-induced cytotoxicity and DNA damage appear to be explained in part by the induction of apoptosis. Results of this study were presented at the 47th Annual Cancer Symposium of The Society of Surgical Oncology, Houston, Texas, March 17–19, 1994     

CCL21/CCR7对T24细胞生物学行为的影响

目的研究CCL21/CCR7对T24细胞迁移、侵袭、增殖以及抗凋亡能力的影响。方法采用不同浓度的CCL21(0、50、100、200ng/ml)作用T24细胞,用Transwell实验研究CCL21对T24细胞的迁移与侵袭能力;用MTT法研究了CCL21对T24细胞的增殖能力的影响,用FCM研究了CCL21对T24细胞抵抗ADM诱导的细胞凋亡,同时用Western blot检测了侵袭相关蛋白MMP-2与MMP-9表达及凋亡相关蛋白Bcl-2与Bax的表达。结果 CCL21作用于T24细胞能增强其迁移和侵袭能力,增强效果依赖于CCL21的浓度,CCL21能增加MMP-2与MMP-9的表达;CCL21还能促进T24细胞的增殖与抗凋亡能力,并能上调Bcl-2蛋白的表达,而抑制Bax蛋白的表达。结论 CCL21/CCR7增强了T24细胞的迁移与侵袭能力,促进了T24细胞的增殖与抗凋亡。