乳腺癌间质成纤维细胞对MDA-MB-231种植肿瘤生长和转移的影响及其机理探讨

目的通过裸鼠接种方式进行体内实验，观察乳腺癌间质成纤维细胞（CAFs）对种植肿瘤生长和转移的影响，并探讨其作用机理。方法选择MDA．MB．231乳腺癌细胞系细胞（简称MDA）、乳腺癌患者的CAFs和癌旁正常乳腺组织的正常成纤维细胞（NFs），结合不同的试剂，包括生理盐水（NS）、基质细胞衍生因子-1（SDF-1）配体拈抗剂1，4，8，11．四氮杂环十四烷（AMD3100，简称AMD），组合成以下6种处理：MDA＋NS、NFs＋NS、MDA＋NFs＋NS、MDA＋NFs＋AMD、MDA＋CAFs＋AMD及MDA＋CAFs＋NS。将36只Bal b／c无胸腺裸小鼠按配伍随机方法分成6组，分别接受上述处理。将细胞悬液接种于裸鼠，接种后饲养46d处死，观察种植肿瘤的大小，有无淋巴结、肺及肝脏转移。采用ELISA法检测6组裸鼠的血浆SDF-1浓度，采用real-timePCR法检测6组裸鼠肿瘤组织中的SDF．1mRNA水平，采用Westernblot法检测6组裸鼠肿瘤组织中SDF-1蛋白的表达水平。结果除NFs＋NS组外，其余5组均有种植肿瘤形成。MDA＋CAFs＋NS组裸鼠的种植肿瘤体积（9．092±2．662）cm3]、血浆SDF．1浓度（75．25±16．23）ng／L]、肿瘤组织中SDF-1 mRNA（中位数为14．714）及其蛋白（中位数为0．673）的表达水平均最高，与其他5组比较差异均有统计学意义（P〈0．050）。6组裸鼠的肝和肺组织均未见转移灶。但MDA＋CAFs＋NS组有4只、MDA＋NS组有2只裸鼠存在腋窝淋巴结转移，其他组均未发现腋窝淋巴结转移。结论乳腺癌CAFs对乳腺癌的生长起促进作用，且可促进肿瘤淋巴结的转移；其作用可能是通过乳腺癌CAFs分泌SDF-1，与其特定的受体即趋化因子受体4（cxcR4）结合来实现的。

Effect and Mechanism of The Breast Cancer Stromal Fibroblasts on Growth and Metastasis of MDA-MB-231 CellsImplanted Tumor

Objective To explore effect and mechanism of the carcinoma associated fibroblasts （CAFs） of breast cancer on growth and metastasis of breast cancer induced in nude mice by inoculation of CAFs and breast Cancer cells. Methods Breast cancer cell line of MDA-MB-231 （abbreviated as MDA）, CAFs, and normal breast tissue fibroblasts （NFs） of the same breast cancer patient were collected, and mixed with normal saline （NS） or SDF-1 ligand blockers of four nitrogen heterocyclic fourteen alka （AMD3100, abbreviated as AMD） for inoculation of nude mice in vivo. According to the different combination, 36 nude mice were randomly divided into 6 groups： MDA＋NS group, NFs＋NS group, MDA＋NFs＋NS group, MDA＋NFs＋AMD group, MDA＋CAFs＋AMD group, and MDA＋CAFs＋NS group. Forty six days after the inoculation and feeding, volume of tumor, metastasis of lymph node, lung or liver were observed. In addition, level of plasma SDF-1 was tested by using ELISA method, and expressions of SDF-1 mRNA and protein intumor specimens were detected by using real-time PCR and Westem blot method respectively. Results Except for NFs ＋ NS group, implanted tumor could be seen in nude mice of other 5 groups. In MDA＋ CAFs ＋NS group, the volume of tumor （9. 092＋2. 662） cm3], level of plasma SDF-1 （75.25＋ 16. 23） ng/L], and expression levels of SDF-1 mRNA （the median level was 14. 714） and protein （the median level was 0. 673）. of tumor tissue were significantly greater or higher than those of the other 5 groups （P〈0. 050）. In addition, lymph node metastasis were found in 4 mice in MDA＋ CAFs＋NS group, and 2 in MDA＋NS group. The tumor metastasis of lung and liver was not found in all nude mice. Conclusion CAFs can promote growth and lymph node metastasis of breast cancer, whose mechanism is related with SDF-1 secreted by CAFs and SDF-1/CXC chemokine receptor 4 （CXCR4）, signal pathway.