现代肿瘤医学
現代腫瘤醫學
현대종류의학
JOURNAL OF MODERN ONCOLOGY
2014年
6期
1249-1253
,共5页
蔡伟%康骅%刘晨%张琳%孙海晨%刘爽%崔叶青%张雁%海涛
蔡偉%康驊%劉晨%張琳%孫海晨%劉爽%崔葉青%張雁%海濤
채위%강화%류신%장림%손해신%류상%최협청%장안%해도
乳腺癌%癌间质成纤维细胞%基质细胞衍生因子-1
乳腺癌%癌間質成纖維細胞%基質細胞衍生因子-1
유선암%암간질성섬유세포%기질세포연생인자-1
breast cancer%carcinoma-associated fibroblasts%SDF-1
目的：通过乳腺癌间质成纤维细胞（ carcinoma-associated fibroblasts，CAFs）与乳腺癌细胞系MDA-MB-231共培养的体外细胞实验及裸鼠接种的在体动物实验，观察CAFs对乳腺癌细胞增殖、凋亡、侵袭和转移活性的影响，并探讨其作用机制。方法：体外实验：分离培养浸润性导管癌组织中CAFs和正常成纤维细胞（ normal fibroblasts，NFs），然后分别与乳腺癌细胞MDA-MB-231体外共培养，采用MTT法、流式细胞仪、Ma-trigel人工模拟基底膜法分别检测乳腺癌细胞的增殖、凋亡、细胞黏附和侵袭能力。动物在体实验：选择乳腺癌细胞系MDA-MB-231、CAFs和NFs，结合生理盐水（NS）、基质细胞衍生因子-1（SDF-1）及其配体拮抗剂AMD3100，组成不同的组别并接种于裸鼠（共6组）。观察肿瘤的大小，有无淋巴结、肺、肝脏转移。留取血标本及肿瘤组织行SDF-1表达水平的检测。结果：MDA-MB-231与CAFs和NFs共培养后乳腺癌细胞的增殖活性显著增强，其中CAFs的作用较NFs更强（P＝0.011）；CAFs组的黏附能力（34.70±4.84个/视野）明显强于NFs组（20.16±3.09个/视野），P＝0.000；而CAFs组的侵袭性（89.0±4.62个/视野）也明显强于NFs组（81.6±6.08个/视野，P＝0.045）。CAFs组中的MDA-MB-231的早期凋亡率（2.9±2.4）较NFs组（5.0±4.2）明显降低（P＝0.026）；MDA231﹢CAFs ﹢NS 组的种植肿瘤平均体积最大（9.092±2.662cm3， P＝0.000）；此外，该组共有4只（66.6％），MDA231﹢NS组有2只（33.3％）存在腋窝淋巴结转移，未见肝肺转移灶。在MDA231﹢CAFs﹢NS组中，血标本 SDF -1值（75.25±16.23pg/ml）、肿瘤组织标本中 SDF -1mRNA值（11.686±8.926）、组织中SDF-1蛋白表达水平（1.006±0.327）均为最高，与其他各组相比均有统计学差异（ P＝0.000）。结论：CAFs可影响乳腺癌肿瘤细胞的生物学特性，具有促进肿瘤细胞增殖，增强其黏附、侵袭及转移能力。其机制可能是通过乳腺癌间质成纤维细胞分泌SDF-1与其特定的受体CXCR4结合这一信号通路来实现的。
目的：通過乳腺癌間質成纖維細胞（ carcinoma-associated fibroblasts，CAFs）與乳腺癌細胞繫MDA-MB-231共培養的體外細胞實驗及裸鼠接種的在體動物實驗，觀察CAFs對乳腺癌細胞增殖、凋亡、侵襲和轉移活性的影響，併探討其作用機製。方法：體外實驗：分離培養浸潤性導管癌組織中CAFs和正常成纖維細胞（ normal fibroblasts，NFs），然後分彆與乳腺癌細胞MDA-MB-231體外共培養，採用MTT法、流式細胞儀、Ma-trigel人工模擬基底膜法分彆檢測乳腺癌細胞的增殖、凋亡、細胞黏附和侵襲能力。動物在體實驗：選擇乳腺癌細胞繫MDA-MB-231、CAFs和NFs，結閤生理鹽水（NS）、基質細胞衍生因子-1（SDF-1）及其配體拮抗劑AMD3100，組成不同的組彆併接種于裸鼠（共6組）。觀察腫瘤的大小，有無淋巴結、肺、肝髒轉移。留取血標本及腫瘤組織行SDF-1錶達水平的檢測。結果：MDA-MB-231與CAFs和NFs共培養後乳腺癌細胞的增殖活性顯著增彊，其中CAFs的作用較NFs更彊（P＝0.011）；CAFs組的黏附能力（34.70±4.84箇/視野）明顯彊于NFs組（20.16±3.09箇/視野），P＝0.000；而CAFs組的侵襲性（89.0±4.62箇/視野）也明顯彊于NFs組（81.6±6.08箇/視野，P＝0.045）。CAFs組中的MDA-MB-231的早期凋亡率（2.9±2.4）較NFs組（5.0±4.2）明顯降低（P＝0.026）；MDA231﹢CAFs ﹢NS 組的種植腫瘤平均體積最大（9.092±2.662cm3， P＝0.000）；此外，該組共有4隻（66.6％），MDA231﹢NS組有2隻（33.3％）存在腋窩淋巴結轉移，未見肝肺轉移竈。在MDA231﹢CAFs﹢NS組中，血標本 SDF -1值（75.25±16.23pg/ml）、腫瘤組織標本中 SDF -1mRNA值（11.686±8.926）、組織中SDF-1蛋白錶達水平（1.006±0.327）均為最高，與其他各組相比均有統計學差異（ P＝0.000）。結論：CAFs可影響乳腺癌腫瘤細胞的生物學特性，具有促進腫瘤細胞增殖，增彊其黏附、侵襲及轉移能力。其機製可能是通過乳腺癌間質成纖維細胞分泌SDF-1與其特定的受體CXCR4結閤這一信號通路來實現的。
목적：통과유선암간질성섬유세포（ carcinoma-associated fibroblasts，CAFs）여유선암세포계MDA-MB-231공배양적체외세포실험급라서접충적재체동물실험，관찰CAFs대유선암세포증식、조망、침습화전이활성적영향，병탐토기작용궤제。방법：체외실험：분리배양침윤성도관암조직중CAFs화정상성섬유세포（ normal fibroblasts，NFs），연후분별여유선암세포MDA-MB-231체외공배양，채용MTT법、류식세포의、Ma-trigel인공모의기저막법분별검측유선암세포적증식、조망、세포점부화침습능력。동물재체실험：선택유선암세포계MDA-MB-231、CAFs화NFs，결합생리염수（NS）、기질세포연생인자-1（SDF-1）급기배체길항제AMD3100，조성불동적조별병접충우라서（공6조）。관찰종류적대소，유무림파결、폐、간장전이。류취혈표본급종류조직행SDF-1표체수평적검측。결과：MDA-MB-231여CAFs화NFs공배양후유선암세포적증식활성현저증강，기중CAFs적작용교NFs경강（P＝0.011）；CAFs조적점부능력（34.70±4.84개/시야）명현강우NFs조（20.16±3.09개/시야），P＝0.000；이CAFs조적침습성（89.0±4.62개/시야）야명현강우NFs조（81.6±6.08개/시야，P＝0.045）。CAFs조중적MDA-MB-231적조기조망솔（2.9±2.4）교NFs조（5.0±4.2）명현강저（P＝0.026）；MDA231﹢CAFs ﹢NS 조적충식종류평균체적최대（9.092±2.662cm3， P＝0.000）；차외，해조공유4지（66.6％），MDA231﹢NS조유2지（33.3％）존재액와림파결전이，미견간폐전이조。재MDA231﹢CAFs﹢NS조중，혈표본 SDF -1치（75.25±16.23pg/ml）、종류조직표본중 SDF -1mRNA치（11.686±8.926）、조직중SDF-1단백표체수평（1.006±0.327）균위최고，여기타각조상비균유통계학차이（ P＝0.000）。결론：CAFs가영향유선암종류세포적생물학특성，구유촉진종류세포증식，증강기점부、침습급전이능력。기궤제가능시통과유선암간질성섬유세포분비SDF-1여기특정적수체CXCR4결합저일신호통로래실현적。
Objective:To explore the effects and mechanism of the breast cancer stromal fibroblasts on the growth, apoptosis,invasion and metastasis of breast cancer cells by the co-culture in vitro and the inoculation of nude mice in vivo. Methods:In vitro:CAFs and NFs were separated from the same patient with infiltrating ductal carcinoma and co-cultured with breast cancer cell line MDA-MB-231 in vitro. The growth,adherence,invasion and apoptosis of breast cancer cells were detected by using the method of MTT,Matrigel membrane or flow cytometer respectively. In vivo:MDA -MB -231 cells,CAFs,or NFs,combined with normal solution( NS ),SDF -1 or its ligand blocker (AMD3100)were inoculated into nude mice(six groups). The tumor size and metastasis of lymph node,lung and liver were observed. Blood SDF-1 levels of nude mice and expression of SDF-1 mRNA and protein in tumor specimens were investigated. Results:The growth of MDA-MB-231 cells were enhanced by co-culturing with CAFs or NFs. CAFs played a more significant effect on breast cells than NFs(P=0. 011). Breast cancer cells co-cultured with CAFs had an obviously stronger adherence(34. 70 ± 4. 84 adhered cells/field)than those with NFs(20. 16 ± 3. 09 ad-hered cells/field),P=0. 000. Breast cancer cells co-cultured with CAFs also had a significantly stronger invasion (89. 0 ± 4. 62 cells/field)than those with NFs(81. 6 ± 6. 08 cells/field),P=0. 045. In flow cytometer assay,MDA-MB-231cells were found less early apoptosis in CAFs group(2. 9 ± 2. 4)than those in NFs group(5. 0 ± 4. 2),P=0. 026. In group of MDA-MB-231 ﹢CAFs﹢NS(group 6),the average tumor volume(9. 092 ± 2. 662 cm3 )was greater than any other groups significantly(P=0. 000). In addition,lymph node metastases were found in four mice in group 6(66. 6%),and 2 in group of MDA-MB-231﹢NS(group 1)(33. 3%). The metastatic tumor was not found in lungs and livers of all nude mice. Blood SDF-1 level(75. 25 ± 16. 23pg/ml),expression level of SDF-1 mRNA (11. 686 ± 8. 926)and SDF-1 protein(1. 006 ± 0. 327)of tumor tissue in group 6 were all higher than the other five groups significantly(P=0. 000). Conclusion:CAFs can promote growth,adherence,invasion and lymph node metas-tasis of breast cancer cells,which mechanism is probably related with SDF-1 secreted by CAFs and SDF-1/ CX-CR4 signal pathway.
