目的：研究TGF-β1对肺癌干细胞和非干细胞增殖的影响，为深入研究肺癌干细胞增殖与分化的分子调控机制奠定基础。方法：利用无血清培养基从人大细胞肺癌组织中分离、培养肿瘤细胞，并建立大细胞肺癌细胞系。以流式细胞术分选表达CD44和(或）CD90的肺癌细胞亚群，通过细胞球形成实验及X射线敏感实验鉴定分选的肺癌干细胞。用流式细胞术及细胞增殖抑制实验检测TGF-β1对肺癌干细胞生长的影响。结果：成功地培养出高纯度的原代大细胞肺癌细胞，其传代次数可达30代以上。应用CD44和CD90共同标记，可将大细胞肺癌细胞分为4亚群，分别是 CD44+CD90+、 CD44+CD90-、CD44++CD90+和CD44++CD90-细胞，其中CD44++CD90+细胞比例为0.4%。相比于其他细胞亚群，有较多的CD44++CD90+细胞为间质细胞样形态，它们形成细胞球的能力最强，对X射线照射的抵抗力也最强。TGF-β1能抑制肺癌细胞的增殖，并促使它们向间质细胞样形态转化。TGF-β1可抑制各亚群肺癌细胞的增殖，但对CD44++CD90+细胞的抑制能力最弱，而对CD44+CD90-细胞的抑制能力最强；培养体系中加入TGF-β1后CD44++CD90+细胞的比例逐步升高，去除TGF-β1后CD44++CD90+细胞的比例又逐渐降低。结论：CD44++CD90+细胞可能是大细胞肺癌的干细胞，TGF-β1能显著抑制肺癌非干细胞的增殖，从而相应地富集了肺癌干细胞。

Objective:To study the effects of TGF-β1 on the growth of cancer stem cells and non-lung cancer stem cells of lung cancer, so as to lay a foundation for studying the molecular regulatory mechanism for proliferation and differentiation of lung cancer stem cells. Methods: Cancer cells were isolated and cultured from human large cell lung cancer specimens using a serum-free medium to establish large cell lung cancer cell line. Biologic characters of lung cancer stem cells were studied by cell sphere formation test and X-ray radiotherapy sensitive assay. The effects of TGF-β1 on the growth of cancer stem cells and non-cancer stem cells of lung cancer were analyzed by flow cytometry and cell growth inhibition test. Results: A large cell lung cancer cell line was established by primary culturing lung cancer tissues. TGF-β1 could inhibit lung cancer cell proliferation and induce them to the shape of mesenchymal cells. When CD44 and CD90 antibodies were co-stained and analyzed by flow cytometry, the lung cancer cells could be divided into 4 populations, i.e. CD44+CD90+, CD44+CD90-, CD44++CD90+ and CD44++CD90- cells. CD44++CD90+cells had the smallest population in the cell line, accounting for 0.4%; they had the strongest cell sphere formation ability and the strongest resistant ability to X-ray radiation among all the 4 populations. TGF-β1 had the weakest suppression effect against CD44++CD90+ cells and the strongest suppression effect against CD44+CD90+ cells. TGF-β1 could gradually increase the ratio of CD44++CD90+ cells when added to the culture medium. After depletion of TGF-β1 from the medium, the ratio of CD44++CD90+ cells gradually decreased again. Conclusion:CD44++CD90+ cells may be the stem cells of large cell lung cancer, and TGF-β1 can enrich lung cancer stem cells in vitro by inhibiting the non-stem cells of lung cancer.

河南省重点科技攻关资助项目（No. 0623033100）；河南省卫生厅重点资助项目（No. 200802015）