大肠癌细胞HCT-15肿瘤干细胞样细胞分离及成瘤性比较

目的:依据人大肠癌细胞HCT-15的表面标志物,分离其中的干细胞亚群。建立裸鼠体内原代大肠癌荷瘤模型,比较各亚群肿瘤体积和质量。方法:利用免疫磁珠分选技术,分离其中CD133/CD44干细胞亚群。分选得到CD133+CD44+、CD133+CD44-亚群分别接种于裸鼠体内,并观察肿瘤大小和质量。结果:CD133+CD44+和CD133+CD44-细胞亚群成功的从HCT-15细胞中分离出来。接种CD133+CD44+细胞的裸鼠形成的肿瘤体积［（2.76±0.22）cm3］和质量［（5.2±0.21）g］明显高于接种CD133+CD44-细胞裸鼠的肿瘤体积与质量［（1.56±0.34）cm3,（3.4±0.18）g］（P＜0.05）。结论:CD44阳性的大肠癌肿瘤干细胞成瘤能力明显高于CD44阴性的大肠癌肿瘤干细胞。     

低剂量节拍化疗对人卵巢癌裸鼠移植瘤中ALDH1表达的影响

目的:探讨顺铂(cisplatin,DDP)最大可耐受剂量化疗(maximum tolerated dose chemotherapy with DDP,MTD-DDP)和DDP低剂量节拍化疗(low-dose metronomic chemotherapy with DDP,LDM-DDP)对人卵巢癌裸鼠移植瘤的作用及其对移植瘤中乙醛脱氢酶1(aldehyde dehydrogenase 1,ALDH1)表达的影响。方法:建立人卵巢癌SKOV3细胞裸鼠皮下移植瘤模型,并进行MTDDDP和LDM-DDP化疗,观察不同化疗模式对移植瘤生长的影响。制备裸鼠移植瘤原代细胞,并应用流式细胞分选术分选ALDH1阳性细胞。应用平板克隆形成实验和裸鼠体内成瘤实验检测ALDH1阳性细胞的克隆形成能力和裸鼠体内成瘤能力,蛋白质印迹法检测ALDH1阳性细胞中Ki-67、乳腺癌耐药蛋白(breast cancer resistant protein,BCRP)和CD133的表达水平。结果:MTD-DDP和LDM-DDP均可抑制裸鼠移植瘤的生长,LDMDDP的抑瘤作用更明显(P<0.001)。MTD-DDP组移植瘤原代细胞中ALDH1阳性细胞所占比率明显高于对照组(以0.9%氯化钠溶液代替DDP),LDM-DDP组移植瘤原代细胞中ALDH1阳性细胞所占比率明显低于对照组,差异均有统计学意义(P值均<0.001)。ALDH1阳性细胞的平板克隆形成能力(P<0.001)和裸鼠体内成瘤能力均高于ALDH1阴性细胞。ALDH1阳性细胞中Ki-67的表达水平低于ALDH1阴性细胞,而BCRP和CD133的表达水平高于ALDH1阴性细胞,差异均有统计学意义(P值均<0.001)。结论:LDM-DDP抑制卵巢癌裸鼠皮下移植瘤生长的作用较强,并可降低ALDH1阳性细胞所占比率。     

喉癌细胞CD44+CD133+高致瘤亚群的生物学特性

目的 筛选喉癌细胞中的高致瘤亚群,探讨其作为肿瘤干细胞的生物学特性.方法 原代培养喉癌细胞,采用流式细胞仪分选CD44+、CD133+、CD44+CD133+、CD44+CD133-细胞,并分别以1×106、1×105、1×104、1×102个/只接种于裸小鼠左腋皮下,观察各亚群成瘤时间、成瘤率、平均瘤重及肿瘤体积,筛选出在极低细胞浓度下的高致瘤亚群.采用Boyden小室体外侵袭实验,检测各亚群细胞的侵袭能力.采用免疫细胞化学染色检测各亚群细胞中干细胞抗原-1(SCA-1)及整合素β1的表达.采用逆转录聚合酶链反应(RT-PCR)和Westem blot法,检测各亚群细胞中Bmi-1基因的表达.结果 CD44+CD133+细胞以1×103个/只接种于裸小鼠,可以成瘤.当各亚群细胞以1×106个/只接种于裸小鼠4周后,CD44+CD133+细胞接种组棵小鼠的肿瘤体积和重量分别为(7726.81±196.93)mm3和(5.51±0.12)g,均大于CD44+、CD44+CD133-及未分选细胞接种组(均P<0.05),而与CD133+细胞接种组裸小鼠的肿瘤体积和重量差异无统计学意义(P>0.05).CD44+CD133+细胞24 h的侵袭细胞数为83.62±1.61,显著高于其他各亚群喉癌细胞.CD44+CD133+细胞高表达整合素β1和SCA-1.Bmi-1 mRNA在CD44+CD133+细胞中的表达水平为0.951±0.112,显著高于CD44+CD133-细胞和未分选的喉癌细胞(0.532±0.214和0.268±0.193,均P<0.01);Bmi-1蛋白在CD133+和CD44+CD133+细胞中高表达,而在CD44+、CD44+CDi33-和未分选的喉癌细胞中仅有少量表达.结论 CD44+CD133+喉癌细胞具有高致瘤性及肿瘤干细胞的生物学特性,可能是喉癌恶性增殖的根源,并有望成为喉癌治疗的新靶点.     

肝癌细胞系Huh-7中CD133阳性细胞亚群的分离及其特性的研究

目的:研究CD133在人肝癌细胞系Huh-7中的表达,初步探讨肝癌中CD133阳性细胞亚群的干细胞特性.方法:流式细胞荧光激活分选技术纯化肝癌细胞系Huh-7中CD133肿瘤细胞,体外培养并观察其增殖、体内成瘤及分化能力.结果:流式细胞仪检测肝癌细胞系Huh-7中有62.3%的细胞CD133呈阳性表达,流式细胞荧光激活分选的CD133肿瘤细胞在无血清培养基中第3、5、7d的吸光度(OD值)分别为0.310、0.362、0.564,均高于相同条件下未分选细胞和CD133阴性细胞;CD133阳性细胞亚群成瘤能力明显强于阴性细胞亚群;CD133阳性细胞亚群在培养体系中的比例逐日下降,至培养的第8天,由第1天的92.3%下降至61.4%.结论:肝癌细胞系Huh-7中CD133阳性细胞亚群比其它细胞亚群具有较强增殖、成瘤和分化能力,是具有肝癌干细胞特性的细胞亚群.     

肝癌干细胞抗体靶向治疗的实验

目的研究抗人肝癌干细胞鼠单抗15B7的生物学特征、体内外功能,探讨靶向肝癌干细胞是否能够有效抑制肝癌移植瘤复发、自发性肺转移以及延长荷瘤小鼠的生存期。方法采用双色免疫荧光、双色流式细胞技术、皮下成瘤实验,检测、鉴定15B7单克隆抗体能够识别肝癌干细胞(hepatocellular carcinomacancer stem cells, HCC-CSC)。从人肝癌细胞系BEL7402中以流式细胞仪分选具有CD133+或ESA+表型的细胞。在此基础上采用CCK-8细胞增殖实验、侵袭实验、迁移实验等检测分析15B7单抗对CD133+表型的细胞增殖、侵袭、迁移的作用以及对细胞周期的影响。裸鼠体内治疗实验研究15B7单抗对BEL7402移植瘤生长的抑制作用。以Western blot方法鉴定该功能性单抗的抗原。结果双色免疫荧光和双色流式检测显示15B7单抗能与HCC-CSC的标志物ESA、CD133共染;流式分选15B7+或ESA+或CD133+的细胞在体外无血清培养条件下有良好的成球生长能力;流式分选的15B7+细胞裸鼠皮下接种1×104个/只,2月可形成肿瘤,以上实验证明15B7单抗是抗肝癌干细胞的单抗。体外功能实验结果显示15B7单抗能够抑制CD133+细胞的增殖、迁移、侵袭,抑制率分别达13.8%、157%和30.9%,同时还能诱导CD133+细胞发生G1期阻滞。体内治疗实验研究结果表明15B7单抗能明显抑制裸鼠肝癌移植瘤生长,抑制率可达60.5%。Western blot显示15B7单抗识别HCC-CSC表达的抗原蛋白相对分子质量约50 kD。结论15B7单抗能明显抑制裸鼠体内人肝移植瘤的生长,为肝癌干细胞的靶向治疗提供有重要应用价值的候选抗体药物。     

肝细胞性肝癌组织CD133+细胞肿瘤干细胞特性的研究

目的 越来越多的研究表明,肝癌细胞系中能检测到肿瘤干细胞(cancer stem cells,CSCs)的存在.本研究旨在探讨从肝细胞性肝癌(hepatocellular carcinoma,HCC)组织样本中分离获得的CD133+细胞是否具有CSCs特性.方法 将2014-02-01-2015-06-30广西医科大学附属肿瘤医院肝胆外科25例手术获得的新鲜HCC组织和对应癌旁组织,采用酶消化法分别消化成单个肝癌细胞和单个肝细胞,利用流式细胞术检测部分单个肝癌细胞和单个肝细胞CD133的表达率.用剩余的单个肝癌细胞进行原代培养,流式细胞术将培养获得的肝癌细胞分选为CD133+和CD133-细胞,通过平板克隆形成实验、肿瘤球形成实验和裸鼠移植瘤形成实验对比分析这两组细胞的CSCs特性.结果 25例HCC组织中CD133的表达率为3.8％～8.3％,平均值为(5.8±1.6)％,而癌旁组织CD133的表达率为0.1％～0.4％,平均值为(0.2±0.1)％,两者比较差异有统计学意义,t=17.12,P＜0.001.CD133+和CD133-细胞的平均克隆率分别为(25.2±0.8)％和(7.6±0.8)％,两者比较差异有统计学意义,t=81.95,P＜0.001.CD133+和CD133-细胞的平均成球率分别为(20.3±0.6)％和(12.5±1.4)％,两者比较差异有统计学意义,t=68.17,P＜0.001.CD133+细胞的裸鼠移植瘤形成能力明显高于CD133-细胞.结论 从HCC组织样本中分离获得的CD133+细胞具有明显的CSCs特性.     

CD133+与CD133-胶质瘤体内再移植性征比较

目的:比较CD133+和CD133-胶质母细胞瘤体内再移植的细胞性征.方法:剥离26只CD133+和2只CD133-胶质母细胞瘤细胞致瘤裸鼠皮下肿瘤组织,分成4份,第1部分肿瘤块制作成冰冻切片,原位凋亡试剂盒检测细胞凋亡率;从第2部分组织块中提取细胞总蛋白,蛋白质印迹法检测转录因子Oct4和Sox2、细胞核增殖抗原(PCNA)、表皮生长因子受体(EGFR)、血管生成因子2(Ang-2)、基质金属蛋白酶2(MMP-2)和MMP-9的表达水平;第3部分组织块剪碎消化成单细胞悬液,流式细胞术检测CD133+肿瘤细胞百分率;取第4部分肿瘤组织进行去胸腺裸鼠皮下再移植,检测成瘤率.结果:CD133+胶质母细胞瘤细胞所致肿瘤组织细胞凋亡率(4.37±0.74)％,低于CD133-胶质母细胞瘤细胞所致肿瘤组织细胞凋亡率(11.14±2.15)％,差异有统计学意义,P＜0.05;CD133+胶质母细胞瘤组织Oct4、Sox2、PCNA、EGFR、Ang2、MMP-2和MMP-9蛋白表达水平相对较高;CD133+和CD133-胶质母细胞瘤细胞所致肿瘤中CD133+细胞百分率分别为(2.47±0.67)％和(0.44±0.14)％,两者致裸鼠皮下成瘤率分别为10/10和4/10.结论:CD133+胶质母细胞瘤细胞所致肿瘤较CD133-胶质母细胞瘤细胞所致肿瘤具有更高的恶性表型,两者体内再移植后成瘤率提高.     

人滑膜肉瘤肿瘤干样细胞分离与鉴定

目的:分离鉴定滑膜肉瘤（SW982）肿瘤干样细胞。方法:体外培养人滑膜肉瘤细胞系SW982,用CD133标记方法检测SW982中肿瘤干细胞的数量,用免疫磁珠分选法进行分选,分选后所得CD133+和CD133-细胞群分别进行以下实验。通过无血清悬浮培养基培养获得CD133+悬浮细胞球,并将其进行重新贴壁、重新成球以检测其自我更新能力。顺铂（cisplatin,CDDP）和阿霉素（doxorubicin,DXR）分别处理CD133+与CD133-贴壁细胞,CD133+贴壁细胞与CD133+悬浮细胞球,MTS检测每两种细胞药物敏感性。Real-time PCR及Western blotting 检测CD133+和CD133-贴壁细胞干细胞相关基因ABCG2、Bmi1、c-Myc、Nanog、Oct3/4、Sox2表达情况。将CD133+和CD133-贴壁细胞分别接种于6~8周雌性BALB/c裸鼠,观察成瘤情况,并将接种后所得肿瘤进行免疫组化,观察接种瘤含CD133情况。结果:SW982细胞系中CD133含量为8.59%,CDDP、DXR对CD133+和CD133-贴壁细胞抑制具有浓度依赖性。CD133+和CD133-细胞均表达干细胞相关基因ABCG2、Bmi1、c-Myc、Nanog、Oct3/4、Sox2。相对于CD133-细胞,CD133+细胞ABCG2、Nanog、Oct3/4、Sox2表达明显升高。CD133+细胞裸鼠体内成瘤率高于CD133-细胞,CD133+接种瘤含CD133较CD133-接种瘤多。结论:SW982细胞系CD133+细胞具有高自我更新能力、高耐药性、高表达干细胞相关基因、高成瘤性,是肿瘤干细胞。     

CD133~+胶质母细胞瘤细胞的分离与恶性行为特征

背景与目的:在恶性胶质瘤中存在一类恶性度极高的前体细胞,它们控制着肿瘤的生长与恶性演进,也是肿瘤复发的根源。本研究对胶质母细胞瘤进行肿瘤干细胞的分离与恶性行为特征分析,旨在验证胶质瘤干细胞的存在,并为未来该领域研究打下基础。方法:充分消化术中切除的胶质母细胞瘤组织块至单细胞悬液,流式细胞术检测CD133+肿瘤细胞百分率,免疫磁珠分选法分离CD133+和CD133-胶质母细胞瘤细胞;免疫荧光法检测两类细胞中巢蛋白(nestin)、胶质纤维酸性蛋白(GFAP)和神经元特异性烯醇化酶(NSE)的表达水平;Transwell双室培养体系检测两类瘤细胞的侵袭能力,血清依赖实验检测两类瘤细胞在低营养条件下的生存能力,软琼脂克隆形成实验检测瘤细胞的克隆形成能力;去胸腺小鼠腹股沟皮下接种瘤细胞,观察成瘤率。结果:流式细胞术检测CD133+肿瘤细胞百分率为(2.31±0.57)%。CD133+细胞高表达nestin,低表达GFAP和NSE;CD133-细胞则相反。与CD133-细胞比较,CD133+细胞具有更高的细胞侵袭能力、低营养耐受能力和克隆形成能力。CD133+细胞在小鼠体内成瘤率为87%(26/30),CD133-细胞为7%(2/30)。结论:CD133+胶质母细胞瘤细胞具有更高的恶性细胞表型,提示其具有高度研究价值。     

肝癌细胞系Huh-7中CD133阳性细胞亚群的分离及其特性的研究

目的：研究CD133在人肝癌细胞系Huh-7中的表达,初步探讨肝癌中CD133阳性细胞亚群的干细胞特性。方法：流式细胞荧光激活分选技术纯化肝癌细胞系Huh-7中CD133肿瘤细胞,体外培养并观察其增殖、体内成瘤及分化能力。结果i流式细胞仪检测肝癌细胞系Huh-7中有62．3％的细胞CD133呈阳性表达,流式细胞荧光激活分选的CD133肿瘤细胞在无血清培养基中第3、5、7d的吸光度（OD值）分别为0．310、0．362、0．564,均高于相同条件下未分选细胞和CD133阴性细胞;CD133阳性细胞亚群成瘤能力明显强于阴性细胞亚群;CD133阳性细胞亚群在培养体系中的比例逐日下降,至培养的第8天,由第1天的92．3％下降至61．4％。结论：肝癌细胞系Huh-7中CD133阳性细胞亚群比其它细胞亚群具有较强增殖、成瘤和分化能力,是具有肝癌干细胞特性的细胞亚群。     

miR-34 is associated with poor prognosis of patients with gallbladder cancer through regulating telomere length in tumor stem cells

miR-34a has been identified as a tumor suppressor in several tumors, but its involvement in gallbladder cancer (GBC) has not been reported. In this study, the miR-34a level and telomere length were measured in 77 gallbladder adenocarcinomas and 36 peritumoral tissues by real-time PCR. Forced miR-34a expression was established by an adenovirus carrying a miR-34a expression cassette. The colony-forming ability of isolated CD44⁺CD133⁺ GBC tumor stem-like cells was measured by matrigel colony assay. The xenograft tumor models were established by inoculating nude mice with CD44⁺CD133⁺cells. Results showed that significantly lower miR-34a expression and longer telomere length were observed in gallbladder adenocarcinoma tissues, which correlated with poor prognosis of GBC patients. Forced overexpression of miR-34a inhibited the colony-forming ability of CD44⁺CD133⁺ GBC tumor stem-like cells in vitro and xenograft tumor growth in vivo. Injection of Ad-miR-34a downregulated PNUTS expression and reduced telomere length in xenograft GBC tumor cells. In conclusion, miR-34a is a tumor suppressor in gallbladder cancer. Both low miR-34a expression and long telomere length are markers for poor prognosis of patients with gallbladder adenocarcinoma. Our study also suggests that the miR-34a gene could be a target for targeting therapy of GBC.     

CD133+ HCC cancer stem cells confer chemoresistance by preferential expression of the Akt/PKB survival pathway

The recent discovery of cancer stem cells (CSCs) has played a pivotal role in changing our view of carcinogenesis and chemotherapy. Based on this concept, CSCs are responsible for the formation and growth of neoplastic tissue and are naturally resistant to chemotherapy, explaining why traditional chemotherapies can initially shrink a tumor but fails to eradicate it in full, allowing eventual recurrence. Recently, we identified a CSC population in hepatocellular carcinoma (HCC) characterized by their CD133 phenotype. However, the molecular mechanism by which it escapes conventional therapies remains unknown. Here, we examined the sensitivity of these cells to chemotherapeutic agents (doxorubicin and fluorouracil) and the possible mechanistic pathway by which resistance may be regulated. Purified CD133+ HCC cells isolated from human HCC cell line and xenograft mouse models survived chemotherapy in increased proportions relative to most tumor cells which lack the CD133 phenotype; the underlying mechanism of which required the preferential expression of survival proteins involved in the Akt/PKB and Bcl-2 pathway. Treatment of CD133+ HCC cells with an AKT1 inhibitor, specific to the Akt/PKB pathway, significantly reduced the expression of the survival proteins that was normally expressed endogenously. In addition, treatment of unsorted HCC cells with both anticancer drugs in vitro significantly enriched the CD133+ subpopulation. In conclusion, our results show that CD133+ HCC cells contribute to chemoresistance through preferential activation of Akt/PKB and Bcl-2 cell survival response. Targeting of this specific survival signaling pathway in CD133+ HCC CSCs may provide a novel therapeutic model for the disease.     

Influence on Biological Behavior of Colon Cancer Stem Cells after RNA Interfering CD133

OBJECTIVE To observe the effects of blocking CD133 gene expression and activation on biological characteristic of the colon cancer stem cells.METHODS CD133+ colon cancer stem cells (CCSCs) were separated from EpCAMhigh CD44+ CCSCs through fluorescenceactivated cell sorting (FACS). The proliferation, the capability of spherical cell formation, neoplasia, and the expression of ABCG2 mRNA of CD133+ CCSCs were observed after the CD133+ CCSCs were infected with LV-CD133shRNA. CD133 negative cells were isolated from EpCAMhigh CD44+ CCSCs with FACS, and the CD133 proteins in CD133- cells were detected with Western blot.RESULTS CD133+ CCSCs were isolated from EpCAMhigh CD44+ CCSCs using FACS, and they accounted for 89.2% in the stem cells. In the experimental group, after the CD133+ CCSCs were knocked down by LV-CD133shRNA RNAi, the growth pattern of the cells in the stem cell culture changed into adherent growth from suspended growth, and couldn't generate spherical cells. Results of MTT assay showed that the CD133+ CCSCs infected with LV-CD133shRNA grew slowly, compared to the cells in the control groups. There was a decrease in the cloning efficiency. The infected cells were transplanted into the BALB/c nude mice. During the observation, no neoplasia was found in the CD133+ cells infected with LV-CD133shRNA. The level of ABCG2 mRNA expression was lowered greatly (P < 0.01). CD133- cells were obtained from the EpCAMhigh CD44+ CCSCs using FACS, in which the expression of CD133 protein was positive.CONCLUSION CD133 retains the biological characteristics of the colon cancer stem cells.     

Epithelial mesenchymal transition correlates with CD24+CD44+ and CD133+ cells in pancreatic cancer

The epithelial-mesenchymal transition (EMT) has been linked to induction of a stem-cell like phenotype, characterized by altered cell surface marker expression and increased tumor formation. The aim of this study was to investigate whether EMT correlates with CD24+CD44+ and CD133+ cells in pancreatic cancer. The morphology of untreated and gemcitabine-treated SW1990 gemcitabine-resistant cells and normal SW1990 cells were compared. NF-κB p65 expression was knocked down using siRNA. Vimentin and E-cadherin expression were analyzed using western blotting, and CD24+CD44+, CD133+ cells were quantified by FACS. Additionally, immunohistochemistry of EMT-associated markers and stem cell-associated markers were performed in 41 cases of human pancreatic ductal adenocarcinoma. In SW1990 gemcitabine-resistant cells, gemcitabine induced a mesenchymal cell phenotype, expression of EMT-related molecular markers and increased CD24+CD44+ and CD133+ cells compared to untreated SW1990 gemcitabine-resistant and SW1990 cells. Knockdown of NF-κB p65 inhibited the ability of gemcitabine to increase the proportion of CD24+CD44+ or CD133+ cells and expression of EMT-related molecular markers. In human pancreatic ductal adenocarcinoma, significant correlations were observed between expression of the EMT-associated markers vimentin and E-cadherin, and stem cell-associated markers CD24, CD133 and CD44. This study demonstrated that EMT correlated with CD24+CD44+ and CD133+ cells in pancreatic cancer. This study also suggests that EMT may induce cancer stem-like cells in pancreatic cancer, with different degrees of EMT probability inducing different proportions of CD24+CD44+ and CD133+ cells.     

小鼠三阴性乳腺癌干样细胞对EMT 发生及其生物学行为的影响*

目的:探讨TA2 小鼠三阴性乳腺癌中乙醛脱氢酶1+（ALDH1+）和CD133+表型的乳腺癌干样细胞在上皮间充质转化（EMT ）发生中的作用,及对其生物学行为的影响。方法:使用流式细胞术分析TA2 小鼠三阴性乳腺癌组织中ALDH1 及CD133 的表达量并分选出具有ALDH1+、ALDH1-、CD133+、CD133-表型的乳腺癌细胞,将其分别接种于TA2 小鼠,根据细胞表型不同设置为AL?DH1+、ALDH1-,CD133+、CD133-组,观察肿瘤生长情况,并制成组织切片,行三阴性乳腺癌中乳腺癌干细胞表面标记物ALDH1、CD133 与EMT 相关蛋白Twist1、E-cadherin 和VE-cadherin的免疫组织化学检测,分析其表达差异。结果:乳腺癌干细胞标记物ALDH1 及CD133 在TA2 小鼠三阴性乳腺癌中表达率分别为31.2% 和6.5% 。ALDH1+、CD133+组肿瘤生成能力明显强于ALDH1-、CD133-组。免疫组织化学结果显示ALDH1、Twist1、VE-cadherin在ALDH1+组的表达明显高于ALDH1-组（均P < 0.05）,E-cadherin在ALDH1+组的表达低于ALDH1-组（P < 0.05）。 CD133、Twist1、VE-cadherin在CD133+组的表达明显高于CD133-组（均P < 0.05）,E-cadherin 在CD133+组的表达低于CD133-组（P < 0.05）。 结论:TA2 小鼠三阴性乳腺癌中ALDH1+和CD133+表型的乳腺癌干样细胞可影响EMT 相关蛋白的表达,并促进三阴性乳腺癌的形成。      

耐顺铂卵巢癌患者腹水中ALDH1阳性肿瘤细胞的生物学特性

目的:探讨卵巢癌顺铂耐药患者腹水中ALDH1阳性肿瘤细胞的生物学特征。方法:从卵巢癌顺铂耐药患者腹水中分离癌细胞,流式分选法得到 ALDH1阳性和ALDH1阴性细胞,荧光定量RT-PCR检测两组细胞的干细胞标志CD44、CD90、CD133及上皮间质化相关标志E-cadherin、N-cadherin、Vimentin及Fibronectin在mRNA水平的表达,MTT法测定细胞增殖曲线和对顺铂的耐药指数,Transwell 小室侵袭实验、克隆形成实验、悬浮成球实验分别观察细胞侵袭能力、增殖分化能力及干细胞的潜能。结果:与ALDH1阴性细胞相比,ALDH1阳性细胞的干细胞标志CD44、CD90、CD133表达升高,上皮化标志E-cadherin降低,间质化标志 N-cadherin、Vimentin表达明显升高。MTT测定生长曲线显示ALDH1阳性细胞增殖明显增快,对顺铂的耐药指数为14.2,ALDH1阳性细胞的穿膜细胞数、克隆形成数以及成球数目显著高于ALDH1 阴性细胞 (P<0.05)。结论:ALDH1阳性细胞具备干细胞特征,其耐药性和侵袭能力增强。