中国肿瘤临床
中國腫瘤臨床
중국종류림상
CHINESE JOURNAL OF CLINICAL ONCOLOGY
2013年
17期
1016-1019
,共4页
王越%任秀宝%李慧%曹水%任宝柱%于文文%张澎%齐静%于津浦
王越%任秀寶%李慧%曹水%任寶柱%于文文%張澎%齊靜%于津浦
왕월%임수보%리혜%조수%임보주%우문문%장팽%제정%우진포
髓系来源抑制细胞%STAT3%T细胞%乳腺癌
髓繫來源抑製細胞%STAT3%T細胞%乳腺癌
수계래원억제세포%STAT3%T세포%유선암
MDSCs%STAT3%T cell%breast cancer
目的：检测STAT3在乳腺癌MDSCs中的磷酸化状态及其对MDSCs免疫抑制活性的影响。方法：收集脐血单个核细胞，免疫磁珠的方法分选其中CD33+细胞，体外与乳腺癌细胞系MDA-MB-231共孵育诱导MDSC生成。Western blot方法分别检测IDO和STATs的表达与磷酸化情况。MDSCs与健康供者外周血T淋巴细胞共孵育，分别加入1-MT和JSI-124来抑制IDO功能和STAT3磷酸化，利用MTT实验和ELISA检测各组T细胞的增殖和细胞因子分泌。结果：Western blot检测发现体外诱导的MDSCs中IDO表达明显增加，同时伴STAT3磷酸化水平升高。加入JSI-124后pSTAT3和IDO表达明显降低。MTT实验中MDSCs明显抑制T细胞增殖，加用IDO特异性抑制剂1-MT或STAT3抑制剂JSI-124后T细胞增殖抑制明显改善（P<0.05），且1-MT组和JSI-124组之间差异无统计学意义。ELISA结果显示MDSCs显著抑制T细胞分泌IFN-γ，促进TGF-β、IL-10释放（P<0.05）。而加用1-MT或JSI-124后，IFN-γ分泌水平升高，而TGF-β、IL-10分泌水平降低（P<0.05），而1-MT组和JSI-124组之间差异无统计学意义。结论：MDSCs中磷酸化STAT3水平升高导致IDO过表达；STAT3的特异性抑制剂JSI-124可以逆转MDSCs对T细胞增殖和Th1类因子分泌的抑制作用。
目的：檢測STAT3在乳腺癌MDSCs中的燐痠化狀態及其對MDSCs免疫抑製活性的影響。方法：收集臍血單箇覈細胞，免疫磁珠的方法分選其中CD33+細胞，體外與乳腺癌細胞繫MDA-MB-231共孵育誘導MDSC生成。Western blot方法分彆檢測IDO和STATs的錶達與燐痠化情況。MDSCs與健康供者外週血T淋巴細胞共孵育，分彆加入1-MT和JSI-124來抑製IDO功能和STAT3燐痠化，利用MTT實驗和ELISA檢測各組T細胞的增殖和細胞因子分泌。結果：Western blot檢測髮現體外誘導的MDSCs中IDO錶達明顯增加，同時伴STAT3燐痠化水平升高。加入JSI-124後pSTAT3和IDO錶達明顯降低。MTT實驗中MDSCs明顯抑製T細胞增殖，加用IDO特異性抑製劑1-MT或STAT3抑製劑JSI-124後T細胞增殖抑製明顯改善（P<0.05），且1-MT組和JSI-124組之間差異無統計學意義。ELISA結果顯示MDSCs顯著抑製T細胞分泌IFN-γ，促進TGF-β、IL-10釋放（P<0.05）。而加用1-MT或JSI-124後，IFN-γ分泌水平升高，而TGF-β、IL-10分泌水平降低（P<0.05），而1-MT組和JSI-124組之間差異無統計學意義。結論：MDSCs中燐痠化STAT3水平升高導緻IDO過錶達；STAT3的特異性抑製劑JSI-124可以逆轉MDSCs對T細胞增殖和Th1類因子分泌的抑製作用。
목적：검측STAT3재유선암MDSCs중적린산화상태급기대MDSCs면역억제활성적영향。방법：수집제혈단개핵세포，면역자주적방법분선기중CD33+세포，체외여유선암세포계MDA-MB-231공부육유도MDSC생성。Western blot방법분별검측IDO화STATs적표체여린산화정황。MDSCs여건강공자외주혈T림파세포공부육，분별가입1-MT화JSI-124래억제IDO공능화STAT3린산화，이용MTT실험화ELISA검측각조T세포적증식화세포인자분비。결과：Western blot검측발현체외유도적MDSCs중IDO표체명현증가，동시반STAT3린산화수평승고。가입JSI-124후pSTAT3화IDO표체명현강저。MTT실험중MDSCs명현억제T세포증식，가용IDO특이성억제제1-MT혹STAT3억제제JSI-124후T세포증식억제명현개선（P<0.05），차1-MT조화JSI-124조지간차이무통계학의의。ELISA결과현시MDSCs현저억제T세포분비IFN-γ，촉진TGF-β、IL-10석방（P<0.05）。이가용1-MT혹JSI-124후，IFN-γ분비수평승고，이TGF-β、IL-10분비수평강저（P<0.05），이1-MT조화JSI-124조지간차이무통계학의의。결론：MDSCs중린산화STAT3수평승고도치IDO과표체；STAT3적특이성억제제JSI-124가이역전MDSCs대T세포증식화Th1류인자분비적억제작용。
Objective:To explore the status of STAT3 phosphorylation in myeloid-derived suppressor cells (MDSCs) of breast cancer and its function in the immunosuppressive effect of MDSCs on proliferation and cytokine secretion of T cells. Methods:CCD33+cells were isolated from healthy umbilical cord, blood-derived, peripheral blood mononuclear cells and were co-cultured with breast cancer cell line MDA-MB-231 in vitro using Transwell plates to induce MDSCs. The untreated CD33+cells were used as con-trols. Idoxuridine (IDO) suppressor expression and STAT3 phosphorylation were examined using Western blot assay. The proliferation and cytokine secretion of T cells, which were co-cultured with MDSCs, were determined by methyl thiazol tetrazolium assay and en-zyme-linked immunosorbent assay. 1-MT and JSI-124 were used to investigate the function of IDO and pSTAT3 in MDSC-mediated T cell immunosuppression. Results:The protein levels of IDO and pSTAT3 in MDSCs were significantly upregulated. MDSCs obviously suppressed T-cell proliferation, which was reversed by 1-MT or JSI-124 (P<0.05). MDSCs could promote TGF-βand IL-10 secretions, but could also remarkably inhibit IFN-γsecretion (P<0.05). After incubation with 1-MT or JSI-124, the increase in TGF-βand IL-10, as well as the decrease in IFN-γ, was significantly reversed. Conclusion:The upregulated pSTAT3 induced the IDO increase in MDSCs. JSI-124 can block MDSC-mediated immunosuppressive effect on T cells in breast cancer.
