Isolation of cancer stem cells from ovarian cancer cell line HO9810 and identification of their biological characteristics

目的 筛选人卵巢癌HO8910细胞株中的干细胞并观察其生物学特性.方法 以常规培养的HO8910细胞为基础(对照组),采用紫杉醇结合无血清培养基悬浮培养法筛选卵巢癌干细胞(干细胞组).对筛选出的干细胞分别采用MTT法检测细胞的增殖情况;体外侵袭实验检测细胞侵袭能力;流式细胞术检测CD24、CD44、CD45、CD133、CD117表达及Hoechst33342染色阳性率;Western blot检测ABCG2、Nanog、Oct4及BCRP等干细胞标志基因的蛋白表达,并对卵巢癌干细胞在裸鼠体内的致瘤性进行检测.结果 (1)在紫杉醇结合无血清培养筛选条件下,部分HO8910细胞能够较好的生长,具有干细胞特性.(2)干细胞组各时间点的细胞增殖能力较对照组明显增强.(3)干细胞组穿膜细胞数较对照组明显增多.(4)干细胞组CD24+、CD44+、CD45+的阳性表达率与对照组比较,差异均无统计学意义;CD133+、CD117+的阳性表达率及Hoechst33342染色阳性率与对照组比较,差异均有统计学意义(P＜0.05).(5)干细胞组ABCG2、Nanog、Oct4、BCRP等干细胞标志基因的蛋白表达水平与对照组比较均明显增强.(6)干细胞组细胞在裸鼠体内致瘤性明显高于对照组.结论 紫杉醇结合无血清培养基悬浮培养法能够筛选出具有干细胞特征的卵巢癌干细胞;卵巢癌干细胞在体外及体内的生物学特性较普通卵巢癌细胞有很大差异.     

干扰核黄素代谢对细胞卵巢癌HO8910细胞顺铂敏感性的影响（英文）北大核心CSCD

目的研究干预核黄素(RIB)代谢途径对卵巢癌HO8910细胞顺铂(DDP)敏感性的影响。方法采用慢病毒包装sh RNA载体干扰RIB转运体2(RFT2)和RIB竞争性抑制剂氯丙嗪(CHL)干预RIB代谢途径。HO8910卵巢癌细胞分为正常细胞对照组、sh RNA对照组、RFT2 sh RNA组、CHL 50μmol·L^(-1)组、DDP 20μmol·L^(-1)组、RFT2 shR NA+DDP组、CHL+DDP组和DDP+RIB 20μmol·L^(-1)组。各组细胞处理48 h后CCK-8方法检测细胞存活;结晶紫染色方法检测克隆形成能力;流式细胞仪检测凋亡、线粒体膜电位、活性氧(ROS)含量和CD44^+CD133^+细胞比例。结果与单独DDP处理组对比,RFT2 sh RNA或CHL联合DDP能明显降低HO8910细胞活性(P<0.01),减少细胞集落形成能力(P<0.01),降低细胞线粒体膜电位(P<0.01),增加细胞ROS含量和细胞凋亡比例(P<0.01),同时减少CD44^+CD133^+细胞比例(P<0.01),RIB可减弱DDP对HO8910细胞的作用(P<0.01)。结论干扰RIB代谢途径能增强DDP对卵巢癌HO8910细胞的杀伤作用,该作用与增强DDP氧化损伤和降低干细胞样肿瘤细胞比例有关;RIB可减弱DDP对HO8910细胞的作用。     

顺铂不同疗程化疗对裸鼠人卵巢癌移植瘤的影响

目的观察顺铂(DDP)不同疗程化疗对裸鼠人卵巢癌移植瘤的影响,并探讨其对乳腺癌耐药蛋白(BCRP)、P-糖蛋白(P-gp)和肿瘤干细胞标记物CD133表达水平的影响。方法用人卵巢癌细胞株SKOV3建立荷人卵巢癌裸鼠皮下移植瘤模型,24只裸鼠随机分为对照组、短疗程化疗组和足疗程化疗组,每组8只。化疗期间观察裸鼠体重和肿瘤生长状况;Western Blot方法检测3组移植瘤组织中BCRP、P-gp、CD133的蛋白表达水平。结果对照组移植瘤体积最大,短疗程化疗组次之,足疗程化疗组最小(P<0.05);足疗程化疗组裸鼠体重低于对照组及短疗程化疗组(P<0.01,P<0.05);足疗程化疗组CD133、BCRP、P-gp蛋白表达水平高于对照组及短疗程化疗组(P<0.01,P<0.05)。结论 DDP足疗程化疗可以更好地抑制荷人卵巢癌裸鼠皮下移植瘤的生长,但残余瘤对化疗抵抗性更强,更难治疗。     

沉默Oct4和Nanog基因抑制胰腺癌干细胞的生物学特征

目的探讨特异短发夹RNA(shRNA)沉默胰腺癌PANC-1肿瘤干细胞(PCSCs)生物学特征的变化及其机制。方法从PANC-1细胞株中分离出CD44+CD24+上皮特异性抗原(ESA)+PCSCs,检测Oct4和Nanog蛋白表达。用慢病毒载体介导的shRNA沉默PCSCs中Oct4和Nanog表达,用荧光定量RT-PCR和Western blot检测干扰效率。通过单克隆形成实验、Transwell小室模型、细胞计数试剂盒8(CCK8)检测Oct4和Nanog对PCSCs增殖、迁移、侵袭和药物敏感性的影响。NOD/SCID小鼠致瘤实验观察Oct4和Nanog对PCSCs致瘤性的影响。结果 PANC-1细胞株中PCSCs占0.1%-0.8%,Oct4和Nanog在PCSCs中呈高表达(P<0.05);慢病毒载体介导的shRNA降低PCSCs中Oct4和Nanog的表达(P<0.05)。沉默Oct4、Nanog后,PCSCs的单克隆形成、增殖、迁移、侵袭能力下降,药物敏感性增强并诱导凋亡(P<0.05)。致瘤实验表明,沉默Oct4和Nanog后,PCSCs致瘤能力下降(P<0.05)。结论 Oct4和Nanog在PCSCs中高表达,慢病毒介导的shRNA可沉默PCSCs中Oct4、Nanog的表达。沉默PCSCs中Oct4和Nanog的表达可抑制PCSCs相关生物学特征,增强其药物敏感性并诱导细胞凋亡。     

紫杉醇对HO8910卵巢癌细胞增殖及其基质金属蛋白酶（MMP—2，MMP—9）表达的影响

目的:研究紫杉醇对肿瘤细胞增殖及基质金属蛋白酶(MMP-2,MMP-9)表达的影响。方法:半定量RT-PCR法检测紫杉醇对体外培养的HO8910卵巢癌细胞MMP-2和MMP-9基因mRNA表达的作用。结果:(1)MTT法检测提示紫杉醇在浓度达100 nmol·L~(-1)以上时,对HO8910细胞生长抑制率>30％(P<0.001),并随浓度增加而升高(r=0.6574,P<0.01);(2)紫杉醇浓度在10 nmol·L~(-1)时MMP-2和MMP-9 mRNA表达降低(P<0.01),其改变呈剂量依赖性下降。结论:紫杉醇在抑制HO8910细胞增殖时,也抑制基质金属蛋白酶MMP-2和MMP-9mRNA的表达,因而,可能产生抑制肿瘤增殖和侵袭转移的作用。     

二甲双胍对骨肉瘤MG-63细胞形态、凋亡和终末分化的影响分析

目的:研究二甲双胍对骨肉瘤MG-63细胞的形态、凋亡和终末分化的影响,分析其与磷酸腺苷激活的蛋白激酶活性及胚胎干细胞标志物Nanog、Oct4表达情况的相关性.方法:选取人骨肉瘤MG63细胞作为研究对象,利用悬浮球生长实验将其诱导为骨肉瘤干细胞,随机将培养的细胞分为实验组和对照组,实验组培养细胞给予二甲双胍处理,观察处理前后骨肉瘤干细胞的形态及分化能力,分析二甲双胍对骨肉瘤干细胞凋亡的影响,并比较干细胞标志物Nanog及Oct4的阳性率,检测二甲双胍作用前后骨肉瘤干细胞中磷酸腺苷激活的蛋白激酶AMPK、雷帕霉素靶蛋白mTOR以及核糖体蛋白S6激酶p70S6K信号通路的表达情况.结果:实验组培养细胞的Nanog阳性率为45.2%,Oct4阳性率为74.7%;对照组培养细胞的Nanog阳性率为73.5%,Oct4阳性率为95.3%;两组培养细胞的Nanog及Oct4的阳性率比较差异均具有统计学意义(P<0.05).实验组培养细胞的磷酸腺苷激活的蛋白激酶AMPK相对表达量明显高于对照组,雷帕霉素靶蛋白mTOR以及核糖体蛋白S6激酶p70S6K的相对表达量明显低于对照组,差异均具有统计学意义(P<0.05).结论:二甲双胍可破坏骨肉瘤细胞的形态,抑制其分化,作用机制可能与Nanog及Oct4表达的下调、AMPK的激活有关.     

紫杉醇联合固体脂质纳米姜黄素对人卵巢癌HO-8910细胞的抑制作用研究

目的:研究紫杉醇(PTX)联合固体脂质纳米姜黄素(SLN-Cur)对人卵巢癌HO-8910细胞体外生长的抑制作用。方法:试验分为4组,即阴性对照(仅含正常生产的HO-8910细胞)、PTX(2.5μmol/L)、SLN-Cur(10μmol/L)、PTX+SLN-Cur(2.5μmol/L+10μmol/L)组。MTT法测定HO-8910细胞增殖抑制率,透射电镜观察HO-8910细胞凋亡超微结构变化,流式细胞仪检测细胞凋亡率与细胞周期分布变化,免疫组化法检测凋亡相关基因蛋白酶的表达。结果:PTX、SLN-Cur、PTX+SLN-Cur均可显著抑制人卵巢癌HO-8910细胞的增殖,其中PTX+SLN-Cur作用最强。与阴性对照组比较,PTX、SLN-Cur、PTX+SLN-Cur组凋亡率显著增加,S期细胞比例逐渐降低,G2/M期细胞比例逐渐增高,MMP-9表达减弱,TIMP-2表达增强,其中PTX+SLN-Cur组表现更显著。结论:SLN-Cur对人卵巢癌HO-8910细胞具有较好的体外抑制效果,与PTX联合化疗有增效减毒作用,其机制可能与下调MMP-9表达及上调TIMP-2表达而诱导细胞凋亡有关。     

顺铂、紫杉醇对人卵巢癌HO-8910细胞端粒酶活性表达的影响

目的　探讨顺铂和紫杉醇对卵巢癌细胞端粒酶活性表达的影响。方法　用TRAP ELISA法分别检测顺铂 (5 μg/ml)和紫杉醇 (10 6M )作用不同时间后卵巢癌HO 8910细胞端粒酶活性改变 ,并以TUNEL法同步检测细胞凋亡。结果　随着药物作用时间的延长 ,HO 8910细胞株的凋亡率逐渐增加 ,而端粒酶活性逐渐降低。在紫杉醇作用 6 0h后 ,细胞凋亡达 (88 30± 1 0 0 ) %时 ,端粒酶才完全丧失活性 ;而顺铂作用 12h后 ,该细胞株的端粒酶活性完全消失时细胞凋亡率仅有(11 4 3± 0 2 3) %。结论　端粒酶活性抑制可能是顺铂诱导卵巢癌细胞凋亡的机制之一 ,临床监测端粒酶活性表达有助于对抗卵巢癌化疗药物的疗效进行客观评价。     

半边旗提取物5F对高转移卵巢癌细胞HO-8910PM侵袭转移的影响

目的研究半边旗提取物5F(以下简称5F)对人高转移卵巢癌细胞HO8910PM转移相关能力的影响及其作用的机制。方法以MTT法检测5F对高转移卵巢癌细胞HO8910PM细胞增殖的影响;以细胞粘附人工重组基底膜实验检测5F对HO8910PM细胞粘附能力的影响;用Transwell小室法检测5F对HO8910PM细胞的侵袭能力和趋化运动能力的影响;Westernblot法检测5F对HO8910PM细胞NFκB(P65)和VEGF蛋白表达的影响。结果50μmol·L-1的5F作用细胞6h后能抑制HO8910PM细胞体外侵袭人工基底膜、趋化运动和粘附能力,抑制率分别为(37.57±0.62)%,(28.42±0.67)%,(46.07±4.49)%;5F能明显下调HO8910PM细胞中NFκB(P65)和VEGF蛋白的表达。结论5F能抑制高转移卵巢癌细胞HO8910PM的侵袭、运动和粘附能力。5F抗肿瘤侵袭转移的作用机制与NFκB(P65)和VEGF蛋白的表达下调有关。     

卵巢癌腹水中肿瘤干/祖细胞的筛选及鉴定

目的:从人卵巢癌腹水细胞中分离肿瘤干/祖细胞并进行鉴定。方法:采用无血清培养法从腹水中分离培养球形体细胞(ASC);用流式细胞仪检测腹水ASC和腹水细胞(AC)中SP(side population)亚群含量;采用PCR和Western blot方法测定两种细胞干/祖细胞相关标志物ABCG2、Oct-4、Nanog基因和蛋白的表达;NOD/SCID小鼠检测其体内致瘤性。结果:腹水球形体细胞和腹水细胞中SP亚群分别为0.9%,0.2%;腹水球形体细胞较腹水细胞高表达干/祖细胞相关标志物Oct-4、ABCG2、Nanog;1×104个球形体细胞就能在NOD/SCID鼠中成瘤,而1×106个腹水细胞也不能成瘤。结论:无血清培养法可以从卵巢癌患者腹水中筛选出肿瘤干/祖细胞。该结果可为今后研究卵巢癌的发生、复发机制及筛选其化疗药物提供简便实用的体外模型。     

MDM2 antagonist nutlin-3a reverses mitoxantrone resistance by inhibiting breast cancer resistance protein mediated drug transport

Breast cancer resistance protein (BCRP; ABCG2), a clinical marker for identifying the side population (SP) cancer stem cell subgroup, affects intestinal absorption, brain penetration, hepatobiliary excretion, and multidrug resistance of many anti-cancer drugs. Nutlin-3a is currently under pre-clinical investigation in a variety of solid tumor and leukemia models as a p53 reactivation agent, and has been recently demonstrated to also have p53 independent actions in cancer cells. In the present study, we first report that nutlin-3a can inhibit the efflux function of BCRP. We observed that although the nutlin-3a IC₅₀ did not differ between BCRP over-expressing and vector control cells, nutlin-3a treatment significantly potentiated the cells to treatment with the BCRP substrate mitoxantrone. Combination index calculations suggested synergism between nutlin-3a and mitoxantrone in cell lines over-expressing BCRP. Upon further investigation, it was confirmed that nutlin-3a increased the intracellular accumulation of BCRP substrates such as mitoxantrone and Hoechst 33342 in cells expressing functional BCRP without altering the expression level or localization of BCRP. Interestingly, nutlin-3b, considered virtually “inactive” in disrupting the MDM2/p53 interaction, reversed Hoechst 33342 efflux with the same potency as nutlin-3a. Intracellular accumulation and bi-directional transport studies using MDCKII cells suggested that nutlin-3a is not a substrate of BCRP. Additionally, an ATPase assay using Sf9 insect cell membranes over-expressing wild-type BCRP indicated that nutlin-3a inhibits BCRP ATPase activity in a dose-dependent fashion. In conclusion, our studies demonstrate that nutlin-3a inhibits BCRP efflux function, which consequently reverses BCRP-related drug resistance.     

Role of breast cancer resistance protein (BCRP/ABCG2) in cancer drug resistance

Since cloning of the ATP-binding cassette (ABC) family member breast cancer resistance protein (BCRP/ABCG2) and its characterization as a multidrug resistance efflux transporter in 1998, BCRP has been the subject of more than two thousand scholarly articles. In normal tissues, BCRP functions as a defense mechanism against toxins and xenobiotics, with expression in the gut, bile canaliculi, placenta, blood-testis and blood-brain barriers facilitating excretion and limiting absorption of potentially toxic substrate molecules, including many cancer chemotherapeutic drugs. BCRP also plays a key role in heme and folate homeostasis, which may help normal cells survive under conditions of hypoxia. BCRP expression appears to be a characteristic of certain normal tissue stem cells termed "side population cells," which are identified on flow cytometric analysis by their ability to exclude Hoechst 33342, a BCRP substrate fluorescent dye. Hence, BCRP expression may contribute to the natural resistance and longevity of these normal stem cells. Malignant tissues can exploit the properties of BCRP to survive hypoxia and to evade exposure to chemotherapeutic drugs. Evidence is mounting that many cancers display subpopulations of stem cells that are responsible for tumor self-renewal. Such stem cells frequently manifest the "side population" phenotype characterized by expression of BCRP and other ABC transporters. Along with other factors, these transporters may contribute to the inherent resistance of these neoplasms and their failure to be cured.     

Coffee induces breast cancer resistance protein expression in Caco-2 cells

Coffee is a beverage that is consumed world-wide on a daily basis and is known to induce a series of metabolic and pharmacological effects, especially in the digestive tract. However, little is known concerning the effects of coffee on transporters in the gastrointestinal tract. To elucidate the effect of coffee on intestinal transporters, we investigated its effect on expression of the breast cancer resistance protein (BCRP/ABCG2) in a human colorectal cancer cell line, Caco-2. Coffee induced BCRP gene expression in Caco-2 cells in a coffee-dose dependent manner. Coffee treatment of Caco-2 cells also increased the level of BCRP protein, which corresponded to induction of gene expression, and also increased cellular efflux activity, as judged by Hoechst33342 accumulation. None of the major constituents of coffee tested could induce BCRP gene expression. The constituent of coffee that mediated this induction was extractable with ethyl acetate and was produced during the roasting process. Dehydromethylepoxyquinomicin (DHMEQ), an inhibitor of nuclear factor (NF)-κB, inhibited coffee-mediated induction of BCRP gene expression, suggesting involvement of NF-κB in this induction. Our data suggest that daily consumption of coffee might induce BCRP expression in the gastrointestinal tract and may affect the bioavailability of BCRP substrates.     

纳米雄黄对MCF-7乳腺癌干细胞的体外抑制作用

目的 研究纳米雄黄对乳腺癌干细胞的体外抑制作用.方法 以人乳腺癌MCF-7亲本细胞为对象,采用无血清培养法培养获得乳腺癌干细胞.以阿霉素(1 mg/L)为阳性对照,以相同质量浓度的水飞雄黄为参照,采用CCK-8法检测纳米雄黄对MCF-7亲本细胞及干细胞增殖的影响;借助悬浮球形成及分化实验、划痕实验、Transwell侵...     

A novel biosensor based on intestinal 3D organoids for detecting the function of BCRP

Breast cancer resistance protein (BCRP), a key drug efflux transporter, significantly affects the therapeutic efficacy of many drugs. Thus, screening specific BCRP inhibitors and distinguishing between substrates and non-substrates of BCRP are valuable in drug discovery and development. This study presents a novel BCRP biosensor based on intestinal 3D organoids for rapid and sensitive detection of BCRP function. First, the crypts were isolated from mouse small intestine, and cultured in advanced DMEM/F12 medium to develop intestinal 3D organoids. Second, immunohistochemical studies demonstrated that BCRP protein in the organoids presented a similar expression and physiologic position to the small intestinal epithelium. Finally, the cultured organoids were treated in BCRP fluorogenic probe substrate Hoechst 33342 with or without BCRP inhibitor Ko143 and YHO-13177. The fluorescence intensity of Hoechst 33342 released from inner of the organoids was detected by microplate reader and the concentrations were calculated. Ko143 and YHO-13177 significantly inhibited the BCRP-mediated Hoechst 33342 transport in the 3D organoids. Consequently, a rapid and efficient biosensor has been successfully established to study BCRP, especially screening BCRP inhibitors in a high-throughput way.     

低频超声联合微泡对比剂通过PI3K/AKT通路对乳腺癌耐药株MCF-7/ADR多药耐药的逆转机制研究

目的 探索低频超声联合微泡（LFUSMB）通过磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）通路对乳腺癌 耐药株MCF-7/阿霉素（ADR）多药耐药的逆转效果及机制。方法 CCK-8法筛选LFUSMB作用时间并测定ADR对 MCF-7/ADR 细胞的半抑制浓度（IC50）。将 MCF-7/ADR 细胞分组为：对照（C）组,采用 MCF 细胞培养基正常培养; ADR组,采用加入ADR（终质量浓度20 mg/L）的MCF细胞培养基培养;LFUSMB+ADR组,采用加入ADR（终质量浓度 20 mg/L）的MCF细胞培养基培养并经LFUSMB处理30 s;LFUSMB+ADR+740 YP（PI3K/AKT通路活化剂）组,采用加 入 ADR（终质量浓度 20 mg/L）和 740 YP（终质量浓度 50 mg/L）的 MCF 细胞培养基培养并经 LFUSMB 处理 30 s。 Annexin V-FIFC/PI 法检测 MCF-7/ADR 细胞凋亡率,Western blot 检测 P 糖蛋白（P-gp）、抗多药耐药蛋白（MRP）1、 MRP2、乳腺癌抗性蛋白（BCRP/ABCG2）、PI3K、AKT、p-PI3K、p-AKT蛋白表达。结果 采用LFUSMB干预30 s进行 后续研究。LFUSMB 30 s组ADR对MCF/ADR细胞的IC50低于未超声处理组,相对耐药逆转倍数约为8.20倍。ADR 组细胞凋亡率较C组增高（P＜0.05）,MRP1、MRP2、P-gp、BCRP/ABCG2、p-PI3K/PI3K和p-AKT/AKT 蛋白水平无明 显变化;LFUSMB+ADR组细胞凋亡率较ADR组增高,MRP1、MRP2、P-gp、BCRP/ABCG2、p-PI3K/PI3K和p-AKT/AKT 蛋白水平显著下调（P＜0.05）。与 LFUSMB+ADR 组比较,LFUSMB+ADR+740 YP 组细胞凋亡率显著降低,MRP1、 MRP2、P-gp、BCRP/ABCG2 蛋白水平显著增高（P＜0.05）。结论 LFUSMB 可通过抑制 PI3K/AKT 通路活化,下调 MRP1等腺苷三磷酸结合盒（ABC）家族蛋白表达,逆转乳腺癌MCF-7/ADR细胞耐药性。     

趋化因子受体CXCR-4在卵巢癌增殖及转移中的作用

目的观察CAOV-3和HO8910卵巢癌细胞株趋化因子受体(CXCR-4)表达及对其增殖、迁移的作用。方法流式细胞术分析CAOV-3及HO8910细胞黏附分子表达;反转录-聚合酶链反应(RT-PCR)检测CXCR-4转录,观察不同浓度基质蛋白衍生因子(SDF)-1α对CAOV-3和HO8910细胞增殖、迁移及其表面黏附分子表达的影响,并采用激光共聚焦显微镜观察经SDF-1α作用后,CAOV-3细胞表面黏附分子分布变化与其移行能力变化的相关性。结果流式细胞术检测提示二株细胞均表达CD44(98.8%和97.7%)、CD29分子(67.8%和73.5%)和CD49d(45.8%vs37.2%),但不表达CD62e(4.2%vs3.2%);CAOV-3细胞尚表达CXCR-4分子,SDF-1α上调CAOV-3细胞CD29分子(67.8%vs93.2%)的表达,尤其是可显著上调CD49e的表达(25.8%vs82.2%,P<0.05),并可促进其细胞增殖和移行,CAOV-3细胞移行与其细胞表面黏附分子重分布有关。结论SDF-1α可促进CAOV-3细胞增殖和移行,上调CD49e和CD29的表达和重分布,在肿瘤的转移中具有重要作用。     

Establishment of a human colorectal cancer cell line P6C with stem cell properties and resistance to chemotherapeutic drugs

Aim:

Cancer stem cells have the capacity to initiate and sustain tumor growth. In this study, we established a CD44⁺ colorectal cancer stem cell line with particular emphasis on its self-renewal capacity, enhanced tumor initiation and drug resistance.

Methods:

Fresh colon cancer and paired normal colon tissues were collected from 13 patients who had not received chemotherapy or radiotherapy prior to surgery. Among the 6 single-cell derived clones, only the P6C cell line was cultured for more than 20 passages in serial culture and formed holoclones with high efficiency, and then the stemness gene expression, colony formation, tumorigenicity and drug sensitivities of the P6C cell line were examined.

Results:

Stemness proteins, including c-Myc, Oct3/4, Nanog, Lgr5, and SOX2, were highly expressed in the P6C cell line. Oct3/4-positive P6C cells mostly generated holoclones through symmetric division, while a small number of P6C cells generated meroclones through asymmetric division. P6C cells stably expressed CD44 and possessed a high capacity to form tumor spheres. A single cell-derived sphere was capable of generating xenograft tumors in nude mice. Compared to SW480 and HCT116 colorectal cancer cells, P6C cells were highly resistant to Camptothecin and 5-fluorouracil, the commonly used chemotherapeutic agents to treat colorectal cancers.

Conclusion:

We established a colorectal cancer stem cell line P6C with a high tumorigenic capacity and the characteristics of normal stem cells. It will benefit the mechanistic studies on cancer stem cells and the development of drugs that specifically target the cancer stem cells.