南方医科大学学报

南方医科大学学报 ›› 2018, Vol. 38 ›› Issue (01): 1-.

• •    下一篇

色素上皮衍生因子通过调控上皮间质转化抑制乳腺癌细胞侵袭和转移

周丹，许鹏程，张敏，于洋，吴爱国   

  • 出版日期:2018-01-20 发布日期:2018-01-20

Pigment epithelium-derived factor inhibits metastasis and invasion of breast cancer via regulation of epithelial-mesenchymal transition

  • Online:2018-01-20 Published:2018-01-20

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摘要： 目的探讨色素上皮衍生因子（PEDF）是否通过作用上皮间质转化（EMT）抑制乳腺癌浸润转移。方法免疫组化方法检测 119例浸润性导管癌组织中PEDF、vimentin、E-cadherin表达情况；构建PEDF-siRNA-vector干扰载体，应用RNA干扰技术阻断 乳腺癌SK-BR-3细胞中PEDF表达，并构建重组腺病毒载体构建（Lentivirus-PEDF-vector），转染SK-BR-3细胞。采用细胞划痕 实验、细胞侵袭实验、Western bolt方法检测SK-BR-3细胞中PEDF表达变化，并釆用Western bolt方法检测乳腺癌细胞上皮性标 志物E-cadherin和间质性标志物vimentin的表达改变，并观察乳腺癌细胞的体外增殖、侵袭、粘附特性的改变。统计分析采用卡 方检验、Fisher精确概率法检验Spearman等级相关检验、配对计数资料检验。结果乳腺浸润性导管癌中PEDF阳性表达率明 显低于正常乳腺组织，PEDF阳性表达与肿瘤大小相关，与E-cadherin表达正相关（r=0.473，P<0.001）, 与vimentin表达呈负相关 （r=-0.412，P<0.001）。乳腺癌SK-BR-3细胞在PEDF条件培养基培养后：细胞形态学发生改变；Transwell侵袭实验表明：细胞 侵袭转移力减弱；PEDF-siRNA增加SK-BR-3细胞的迁移与侵袭，而Lentivirus-PEDF-vector抑制SK-BR-3细胞的侵袭与迁移 能力。Western blot结果显示：细胞中E-cadherin表达明显减少（P<0.05），vimentin表达明显增多（P<0.05）。结论PEDF基因与 乳腺癌的浸润转移过程密切相关，PEDF可作用SK-BR-3细胞发生EMT进而抑制乳腺癌的浸润转移。

Abstract: Objective To investigate whether pigment epithelium-derived factor (PEDF) inhibits invasion and metastasis of breast cancer through regulation of epithelial-mesenchymal transition. Methods The expressions of PEDF, vimentin, and E-cadherin were detected in 119 breast cancer tissues using immunohistochemistry. SK-BR-3 breast cancer cell models of PEDF knockdown and PEDF overexpression were established by transfecting the cells with a PEDF-siRNA vector and a lentivirus- PEDF vector, respectively. Western blotting was used to detect the changes in the expressions of PEDF, vimentin, and E-cadherin in the cells, and the cell invasion and migration ability was assessed using scratch wound healing assay and transwell migration assay. Results PEDF positivity rate was significantly lowered in breast cancer tissues compared with the adjacent tissues. PEDF was positively correlated with the tumor size and the expression level of E-cadherin (r=0.473, P<0.001), but was negatively correlated with vimentin expression (r=-0.412, P<0.001). Transwell invasion experiment showed that PEDF interference enhanced the cell invasion and metastasis, while PEDF overexpression inhibited the invasion and migration of SKBR- 3 cells. Western blotting showed that PEDF knockdown significantly decreased the expression of E-cadherin (P<0.05) and increased vimentin expression in the cells (P<0.05). Conclusion PEDF is closely related with the metastasis of breast cancer cells through regulation of epithelial-mesenchymal transition.