趋化因子受体CXCR4及其配体CXCL12在肝癌侵袭转移中的作用及机制
目的 探讨趋化因子受体 CXCR4 及其配体 CXCL12 （CXCL12/CXCR4） 在原发性肝癌侵袭转移中的作用及机制.方法 分别采用 Westernblot、 免疫组化和 Real-timePCR 等方法检测 60 例肝癌及对应癌旁组织标本中CXCL12/CXCR4 蛋白及 mRNA 表达水平.常规培养 4 种肝癌细胞 （Huh7、 MHCC97h、 HepG2、 Hep3B） 和正常肝细胞（7702）, Real-timePCR 检测上述细胞中 CXCL12、 CXCR4 mRNA 的表达, 筛选合适的实验细胞.将 CXCR4 干扰质粒 （sh-CXCR4） 和对应空载体 （sh-c...
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| Published in | 天津医药 Vol. 46; no. 1; pp. 20 - 26 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
南昌大学第二附属医院肝胆外科,330000
2018
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0253-9896 |
| DOI | 10.11958/20170729 |
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| Abstract | 目的 探讨趋化因子受体 CXCR4 及其配体 CXCL12 （CXCL12/CXCR4） 在原发性肝癌侵袭转移中的作用及机制.方法 分别采用 Westernblot、 免疫组化和 Real-timePCR 等方法检测 60 例肝癌及对应癌旁组织标本中CXCL12/CXCR4 蛋白及 mRNA 表达水平.常规培养 4 种肝癌细胞 （Huh7、 MHCC97h、 HepG2、 Hep3B） 和正常肝细胞（7702）, Real-timePCR 检测上述细胞中 CXCL12、 CXCR4 mRNA 的表达, 筛选合适的实验细胞.将 CXCR4 干扰质粒 （sh-CXCR4） 和对应空载体 （sh-control） 分别转染至 MHCC97h 构建稳转细胞系.Transwell 侵袭实验、 细胞划痕实验、 MTT 实验分别检测 2 组细胞的侵袭、 迁移和增殖能力.取稳定表达的 sh-control 和 sh-CXCR4 MHCC97h 细胞,接种至 6 只裸鼠皮下, 观察瘤体生长情况.Westernblot 检测 sh-control 和 sh-CXCR4 MHCC97h 细胞及对应裸鼠移植瘤中血管内皮生长因子-C （VEGF-C） 的表达, 同时对转染 CXCR 过表达质粒的 MHCC97h 中 VEGF-C 的表达水平进行检测.结果 （1） Westernblot、 免疫组化和 Real-timePCR 的结果均证实肝癌组织中 CXCL12/CXCR4 蛋白及mRNA 表达水平均高于癌旁组织.（2） Huh7、 MHCC97h、 HepG2、 Hep3B 细胞中 CXCL12/CXCR4 mRNA 表达水平均高于 7702 细胞, 选取 MHCC97h 为实验细胞.转染 sh-CXCR4 的 MHCC97h 细胞的侵袭、 迁移和增殖能力均明显低于sh-control 组, 同时接种含 sh-CXCR4 的 MHCC97h 细胞的裸鼠移植瘤的生长速度也明显小于 sh-control 组.（3） 体外和体内实验均证实 sh-CXCR4 组的 VEGF-C 表达水平均低于 sh-control 组, 而过表达 CXCR4 后, VEGF-C 的蛋白表达明显上调.结论 CXCL12/CXCR4 在原发性癌组织和肝癌细胞中呈现高表达, CXCL12/CXCR4 可能通过调控VEGF-C 蛋白表达从而抑制肝癌细胞的增殖、 侵袭和 |
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| AbstractList | R735.7; 目的 探讨趋化因子受体CXCR4及其配体CXCL12(CXCL12/CXCR4)在原发性肝癌侵袭转移中的作用及机制.方法 分别采用Western blot、免疫组化和Real-time PCR等方法检测60例肝癌及对应癌旁组织标本中CXCL12/CXCR4蛋白及mRNA表达水平.常规培养4种肝癌细胞(Huh7、MHCC97h、HepG2、Hep3B)和正常肝细胞(7702),Real-time PCR检测上述细胞中CXCL12、CXCR4 mRNA的表达,筛选合适的实验细胞.将CXCR4干扰质粒(sh-CXCR4)和对应空载体(sh-control)分别转染至MHCC97h构建稳转细胞系.Transwell侵袭实验、细胞划痕实验、MTT实验分别检测2组细胞的侵袭、迁移和增殖能力.取稳定表达的sh-control和sh-CXCR4 MHCC97h细胞,接种至6只裸鼠皮下,观察瘤体生长情况.Western blot检测sh-control和sh-CXCR4 MHCC97h细胞及对应裸鼠移植瘤中血管内皮生长因子-C(VEGF-C)的表达,同时对转染CXCR过表达质粒的MHCC97h中VEGF-C的表达水平进行检测.结果 (1)Western blot、免疫组化和Real-time PCR的结果均证实肝癌组织中CXCL12/CXCR4蛋白及mRNA表达水平均高于癌旁组织.(2)Huh7、MHCC97h、HepG2、Hep3B细胞中CXCL12/CXCR4 mRNA表达水平均高于7702细胞,选取MHCC97h为实验细胞.转染sh-CXCR4的MHCC97h细胞的侵袭、迁移和增殖能力均明显低于sh-control组,同时接种含sh-CXCR4的MHCC97h细胞的裸鼠移植瘤的生长速度也明显小于sh-control组.(3)体外和体内实验均证实sh-CXCR4组的VEGF-C表达水平均低于sh-control组,而过表达CXCR4后,VEGF-C的蛋白表达明显上调.结论 CXCL12/CXCR4在原发性癌组织和肝癌细胞中呈现高表达,CXCL12/CXCR4可能通过调控VEGF-C蛋白表达从而抑制肝癌细胞的增殖、侵袭和转移. 目的 探讨趋化因子受体 CXCR4 及其配体 CXCL12 （CXCL12/CXCR4） 在原发性肝癌侵袭转移中的作用及机制.方法 分别采用 Westernblot、 免疫组化和 Real-timePCR 等方法检测 60 例肝癌及对应癌旁组织标本中CXCL12/CXCR4 蛋白及 mRNA 表达水平.常规培养 4 种肝癌细胞 （Huh7、 MHCC97h、 HepG2、 Hep3B） 和正常肝细胞（7702）, Real-timePCR 检测上述细胞中 CXCL12、 CXCR4 mRNA 的表达, 筛选合适的实验细胞.将 CXCR4 干扰质粒 （sh-CXCR4） 和对应空载体 （sh-control） 分别转染至 MHCC97h 构建稳转细胞系.Transwell 侵袭实验、 细胞划痕实验、 MTT 实验分别检测 2 组细胞的侵袭、 迁移和增殖能力.取稳定表达的 sh-control 和 sh-CXCR4 MHCC97h 细胞,接种至 6 只裸鼠皮下, 观察瘤体生长情况.Westernblot 检测 sh-control 和 sh-CXCR4 MHCC97h 细胞及对应裸鼠移植瘤中血管内皮生长因子-C （VEGF-C） 的表达, 同时对转染 CXCR 过表达质粒的 MHCC97h 中 VEGF-C 的表达水平进行检测.结果 （1） Westernblot、 免疫组化和 Real-timePCR 的结果均证实肝癌组织中 CXCL12/CXCR4 蛋白及mRNA 表达水平均高于癌旁组织.（2） Huh7、 MHCC97h、 HepG2、 Hep3B 细胞中 CXCL12/CXCR4 mRNA 表达水平均高于 7702 细胞, 选取 MHCC97h 为实验细胞.转染 sh-CXCR4 的 MHCC97h 细胞的侵袭、 迁移和增殖能力均明显低于sh-control 组, 同时接种含 sh-CXCR4 的 MHCC97h 细胞的裸鼠移植瘤的生长速度也明显小于 sh-control 组.（3） 体外和体内实验均证实 sh-CXCR4 组的 VEGF-C 表达水平均低于 sh-control 组, 而过表达 CXCR4 后, VEGF-C 的蛋白表达明显上调.结论 CXCL12/CXCR4 在原发性癌组织和肝癌细胞中呈现高表达, CXCL12/CXCR4 可能通过调控VEGF-C 蛋白表达从而抑制肝癌细胞的增殖、 侵袭和 |
| Abstract_FL | Objective To investigate the mechanism and function of chemokine receptor CXCR4 and its ligand CXCL12 (CXCL12 / CXCR4) in primary hepatocellular carcinoma (PHC). Methods Western blot assay, immunohistochemistry and Real-time PCR were used to detect the protein and mRNA expressions of CXCL12/CXCR4 in 60 PHC and corresponding paracancerous tissue samples. Four kinds of hepatoma cells (Huh7, MHCC97h, HepG2 and Hep3B) and normal hepatocytes (7702) were routinely cultured, and then real-time PCR was performed to detect the mRNA expressions of CXCL12/CXCR4 in these cells to screen suitable experimental cells. CXCR4 interference plasmid (sh-CXCR4) and corresponding empty vector (sh-control) were transfected into MHCC97h to construct stable transfected cell lines. The ability of invasion, migration, and proliferation of the 2 groups of cells were detected by Tanswell invasion experiment, cell scratch test and MTT test. The stably expressed sh-control and sh-CXCR4 MHCC97h cells were taken into the subcutaneous of six nude mice, and the growth of the tumor was observed. Western blot assay was used to detect the expressions of vascular endothelial growth factor-C (VEGF-C) in sh-control and sh-CXCR4 MHCC97h cell lines and corresponding xenografts in nude mice, as the same in MHCC97h, which was transfected with CXCR overexpressed plasmid. Results (1) The results of Western blot assay, immunohistochemistry and Real-time PCR showed that the expressions of CXCL12/CXCR4 protein and mRNA were significantly higher in liver cancer tissues than those of paracancerous tissues. (2) The expression levels of CXCL12/CXCR4 mRNA were higher in Huh7, MHCC97h, HepG2 and Hep3B cells than those of 7702 cells. MHCC97h was selected as the experimental cells. The ability of invasion, migration and proliferation of MHCC97h cells transfected with sh-CXCR4 were significantly lower than those of sh-control group. Meanwhile, the growth rate of nude mice transplanted with sh-CXCR4 MHCC97h cells was also significantly lower than that of sh-control group. (3) Both in vitro and in vivo experiments showed that the expression of VEGF-C was lower in sh-CXCR4 group than that in sh-control group, and the expression of VEGF-C was obviously up-regulated after overexpression of CXCR4. Conclusion High expressions of CXCL12/CXCR4 are found in primary cancer tissues and hepatoma cells. CXCL12/CXCR4 may inhibit the proliferation and invasion of hepatoma cells by regulating the expression of VEGF-C protein. |
| Author | 张引;余力信;张发鹏;余晓雯;王文龙;罗志强 |
| AuthorAffiliation | 南昌大学第二附属医院肝胆外科,330000 |
| AuthorAffiliation_xml | – name: 南昌大学第二附属医院肝胆外科,330000 |
| Author_FL | LUO Zhi-qiang ZHANG Yin WANG Wen-long YU Li-xin ZHANG Fa-peng YU Xiao-wen |
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| Keywords | receptors,CXCR4 动物实验 cultured cell proliferation 受体,CXCR4 liver neoplasms 趋化因子CXCL12 肿瘤细胞,培养的 chemokine CXCL12 肝肿瘤 血管内皮生长因子受体 animal experimentation vascular endothelial growth factor receptor tumor cells 细胞增殖 |
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| Snippet | 目的 探讨趋化因子受体 CXCR4 及其配体 CXCL12 （CXCL12/CXCR4） 在原发性肝癌侵袭转移中的作用及机制.方法 分别采用 Westernblot、 免疫组化和 Real-timePCR 等方法检测... R735.7; 目的 探讨趋化因子受体CXCR4及其配体CXCL12(CXCL12/CXCR4)在原发性肝癌侵袭转移中的作用及机制.方法 分别采用Western blot、免疫组化和Real-time PCR等方法检测60... |
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| SubjectTerms | CXCR4;肿瘤细胞 培养的;细胞增殖;动物实验;血管内皮生长因子受体 肝肿瘤;趋化因子CXCL12;受体 |
| Title | 趋化因子受体CXCR4及其配体CXCL12在肝癌侵袭转移中的作用及机制 |
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