中国肿瘤临床
中國腫瘤臨床
중국종류림상
CHINESE JOURNAL OF CLINICAL ONCOLOGY
2014年
22期
1417-1421
,共5页
朱玉芬%任美敬%谷峰%付丽
硃玉芬%任美敬%穀峰%付麗
주옥분%임미경%곡봉%부려
乳腺癌%乳腺癌干细胞%肿瘤微环境%无血清培养%微球体
乳腺癌%乳腺癌榦細胞%腫瘤微環境%無血清培養%微毬體
유선암%유선암간세포%종류미배경%무혈청배양%미구체
breast cancer%breast cancer stem cells%tumor microenvironment%serum-free culture%mammospheres
目的：探讨肿瘤微环境在乳腺癌干细胞（breast cancer stem cells，BCSCs）培养鉴定及分化过程中的影响及意义。方法：采用无血清培养液PCM-2及成纤维细胞上清液对乳腺癌细胞及MCF-7细胞进行原代培养。观察乳腺癌细胞微球体形成状况，MTT比色法检测乳腺癌细胞的增殖能力，免疫细胞化学方法检测乳腺癌干细胞标记物及上皮间质标记物的表达，并通过RT-PCR进行验证。结果：无血清培养液PCM-2培养的原代细胞微球体的直径大于成纤维细胞上清液的培养（t=4.996，P=0.002），且原代细胞中ALDH1（aldehyde dehydrogenase 1）的表达率高于后者。成纤维细胞上清液培养的细胞生长速度较无血清培养液PCM-2快，差异具有统计学意义（P=0.004）。RT-PCR检测发现无血清培养液PCM-2培养的原代细胞中ALDH1表达上调，E-cadherin、Vimentin表达下调。结论：在乳腺癌原代细胞和MCF-7细胞中可以采用无血清悬浮培养方法富集BCSCs样微球体，成纤维细胞上清液能够促进BCSCs样微球体的增殖与分化。提示乳腺肿瘤微环境在乳腺癌细胞的生长增殖过程中发挥了至关重要的作用。
目的：探討腫瘤微環境在乳腺癌榦細胞（breast cancer stem cells，BCSCs）培養鑒定及分化過程中的影響及意義。方法：採用無血清培養液PCM-2及成纖維細胞上清液對乳腺癌細胞及MCF-7細胞進行原代培養。觀察乳腺癌細胞微毬體形成狀況，MTT比色法檢測乳腺癌細胞的增殖能力，免疫細胞化學方法檢測乳腺癌榦細胞標記物及上皮間質標記物的錶達，併通過RT-PCR進行驗證。結果：無血清培養液PCM-2培養的原代細胞微毬體的直徑大于成纖維細胞上清液的培養（t=4.996，P=0.002），且原代細胞中ALDH1（aldehyde dehydrogenase 1）的錶達率高于後者。成纖維細胞上清液培養的細胞生長速度較無血清培養液PCM-2快，差異具有統計學意義（P=0.004）。RT-PCR檢測髮現無血清培養液PCM-2培養的原代細胞中ALDH1錶達上調，E-cadherin、Vimentin錶達下調。結論：在乳腺癌原代細胞和MCF-7細胞中可以採用無血清懸浮培養方法富集BCSCs樣微毬體，成纖維細胞上清液能夠促進BCSCs樣微毬體的增殖與分化。提示乳腺腫瘤微環境在乳腺癌細胞的生長增殖過程中髮揮瞭至關重要的作用。
목적：탐토종류미배경재유선암간세포（breast cancer stem cells，BCSCs）배양감정급분화과정중적영향급의의。방법：채용무혈청배양액PCM-2급성섬유세포상청액대유선암세포급MCF-7세포진행원대배양。관찰유선암세포미구체형성상황，MTT비색법검측유선암세포적증식능력，면역세포화학방법검측유선암간세포표기물급상피간질표기물적표체，병통과RT-PCR진행험증。결과：무혈청배양액PCM-2배양적원대세포미구체적직경대우성섬유세포상청액적배양（t=4.996，P=0.002），차원대세포중ALDH1（aldehyde dehydrogenase 1）적표체솔고우후자。성섬유세포상청액배양적세포생장속도교무혈청배양액PCM-2쾌，차이구유통계학의의（P=0.004）。RT-PCR검측발현무혈청배양액PCM-2배양적원대세포중ALDH1표체상조，E-cadherin、Vimentin표체하조。결론：재유선암원대세포화MCF-7세포중가이채용무혈청현부배양방법부집BCSCs양미구체，성섬유세포상청액능구촉진BCSCs양미구체적증식여분화。제시유선종류미배경재유선암세포적생장증식과정중발휘료지관중요적작용。
Objective:To investigate the influence and significance of tumor microenvironment in breast cancer stem-cell culture and identification. Methods: Cells isolated from primary breast cancer tissues were cultured in vitro in a serum-free medium PCM-2 and in the supernatant of cultured fibroblasts. The MCF-7 breast cancer cell line was used as the control group. The status of the micro-spheres was observed, and the proliferative capacity of the cells was detected by methyl thiazolyl tetrazolium assay. The expression of stem cell and epithelial-mesenchymal markers were detected by real-time reverse transcription polymerase chain reaction. Results:The diameter of microspheres in PCM-2 gradually increased with prolonged incubation time (t=4.996, P=0.002). The cells in the superna-tant of cultured fibroblasts increased daily and mostly exhibited a spindle cell growth. The growth rate of primary breast cancer cells was faster in the supernatant of cultured fibroblasts than in PCM-2 (P=0.004). Compared with the case of cells in the supernatant of cul-tured fibroblasts, aldehyde dehydrogenase 1 was upregulated in the primary breast cancer cells cultured in serum-free medium PCM-2, whereas E-cadherin and Vimentin were downregulated. Conclusion:Serum-free culture can be one of the best methods for enriching breast cancer stem cell-like mammospheres. The tumor micro-environment serves a vital function in the growth and development of tu-mor cells and in the evolution of breast cancer.
