Prostate tumor neuroendocrine differentiation via EMT: The road less traveled

The long-standing challenge in the treatment of prostate cancer is to overcome therapeutic resistance during progression to lethal disease.Aberrant transforming-growth factor-b(TGF-b)signaling accelerates prostate tumor progression in a transgenic mouse model via effects on epithelial-mesenchymal transition(EMT),and neuroendocrine differentiation driving tumor progression to castration-resistant prostate cancer(CRPC).Neuroendocrine prostate cancer(NEPC)is highly aggressive exhibiting reactivation of developmental programs associated with EMT induction and stem cell-like characteristics.The androgen receptor(AR)is a critical driver of tumor progression as well as therapeutic response in patients with metastatic CRPC.The signaling interactions between the TGF-β mechanistic network and AR axis impact the EMT phenotypic conversions,and perturbation of epithelial homeostasis via EMT renders a critical venue for epithelial derived tumors to become invasive,acquire the neuroendocrine phenotype,and rapidly metastasize.Combinations of microtubule targeting taxane chemotherapy and androgen/AR targeting therapies have survival benefits in CRPC patients,but therapeutic resistance invariability develops,leading to mortality.Compelling evidence from our group recently demonstrated that chemotherapy(cabazitaxel,second line taxane chemotherapy),or TGF-β receptor signaling targeted therapy,caused reversion of EMT to mesenchymal-epithelial transition and tumor re-differentiation,in in vitro and in vivo prostate cancer models.In this review,we discuss the functional contribution of EMT dynamic changes to the development of the neuroendocrine phenotypedthe newly characterized pathological feature of prostate tumors in the context of the tumor microenvironment-navigated cell lineage changes and the role of this neuroendocrine phenotype in metastatic progression and therapeutic resistance.

Isolation and identification of adipose-derived stromal/stem cells from breast cancer patients after exposure neoadjuvant chemotherapy

BACKGROUND With recent research advances,adipose-derived stromal/stem cells(ASCs)have been demonstrated to facilitate the survival of fat grafts and thus are increasingly used for reconstructive procedures following surgery for breast cancer.Unfortunately,in patients,following radiation and chemotherapy for breast cancer suggest that these cancer treatment therapies may limit stem cell cellular functions important for soft tissue wound healing.For clinical translation to patients that have undergone cancer treatment,it is necessary to understand the effects of these therapies on the ASC's ability to improve fat graft survival in clinical practice.AIM To investigate whether the impact on ASCs function capacity and recovery in cancer patients may be due to the chemotherapy.METHODS ASCs were isolated from the cancerous side and noncancerous side of the breast from the same patients with receiving neoadjuvant chemotherapy(NAC)or notreceiving NAC.ASCs were in vitro treated with 5-fluorouracil(5-FU),doxorubicin(DXR),and cyclophosphamide(Cytoxan)at various concentrations.The stem cells yield,cell viability,and proliferation rates were measured by growth curves and MTT assays.Differentiation capacity for adipogenesis was determined by qPCR analysis of the specific gene markers and histological staining.RESULTS No significant differences were observed between the yield of ASCs in patients receiving NAC treatment and not-receiving NAC.ASCs yield from the cancerous side of the breast showed lower than the noncancerous side of the breast in both patients receiving NAC and not-receiving NAC.The proliferation rates of ASCs from patients didn’t differ much before and after NAC upon in vitro culture,and these cells appeared to retain the capacity to acquire adipocyte traits simile to the ASCs from patients not-receiving NAC.After cessation and washout of the drugs for another a week of culturing,ASCs showed a slow recovery of cell growth capacity in 5-FU-treated groups but was not observed in ASCs treated with DXR groups.CONCLUSION Neoadjuvant therapies do not affect the functioning capacity of ASCs.ASCs may hold great potential to serve as a cell source for fat grafting and reconstruction in patients undergoing chemotherapy.

Biological characteristics of breast carcinomas with neuroendocrine cell differentiation

Background The aim of this study was to investigate DNA content and expression of c-erbB-2, PS2, and prostate-specific antigen (PSA) proteins in breast carcinomas with neuroendocrine (NE) cell differentiation.Methods Chromogranin, c-erbB-2, PS2, and PSA in 131 samples of breast cancer were detected immunohistochemically. Classic Feulgen staining image analysis techniques were used to quantify DNA content in 81 of the breast cancer samples.Results The c-erbB-2 positive rate in breast carcinoma samples containing neuroendocrine cells was 37.5% and the rate of high expression of c-erbB-2 (++ or +++) was 33.3%, both significantly lower than that in breast carcinomas without neuroendocrine cells (62.6% and 68.7%, respectively, P<0.05). The rates of positive PS2 and PSA expression in breast carcinoma samples containing neuroendocrine cells were 72.2% and 55.0%, respectively, both significantly higher than that in breast carcinoma samples without neuroendocrine cells (45.0% and 16.4%, respectively, P<0.05). In NE (+) samples, the integral optical density, DNA index, DNA stemline peak, > 5c aneuploidy cells, and rate of aneuploidy among cells were all lower than that in NE (-) breast carcinomas (P<0.01). In NE (+) grade I or II breast carcinomas, these indices were also all lower than that in the NE (-) breast carcinoma samples (P<0.01).Conclusion Breast carcinomas with neuroendocrine differentiation have a lower rate of malignancy. Neuroendocrine differentiation could serve as a prognostic marker in clinical practice.

Role of surgical resection for non-colorectal non-neuroendocrine liver metastases

It is widely accepted that the indications for hepatectomy in colorectal cancer liver metastases and liver metastases of neuro-endocrine tumors result in relatively better prognoses, whereas, the indications and prognoses of hepatectomy for non-colorectal non-neuroendocrine liver metastases(NCNNLM) remain controversial owing to the limited number of cases and the heterogeneity of the primary diseases. There have been many publications on NCNNLM; however, its background heterogeneity makes it difficult to reach a specific conclusion. This heterogeneous disease group should be discussed in the order from its general to specific aspect. The present review paper describes the general prognosis and risk factors associated with NCNNLM while specifically focusing on the liver metastases of each primary disease. A multidisciplinary approach that takes into consideration appropriate timing for hepatectomy combined with chemotherapy may prolong survival and/or contribute to the improvement of the quality of life while giving respite from systemic chemotherapy.

奥氮平对乳腺癌相关巨噬细胞增殖与分化功能的影响

目的探讨奥氮平对乳腺癌细胞共培养体系中人急性髓系白血病细胞系THP-1细胞增殖和分化功能的影响。方法体外培养THP-1细胞、人乳腺癌MDA-MB-231细胞,建立THP-1与MDA-MB-231共培养组,用CCK-8法检测奥氮平处理后的THP-1单独组、共培养组中THP-1的活性变化;用qRT-PCR法检测奥氮平对佛波酯诱导的THP-1单独组、共培养组中THP-1表面分子CD68、CD80、CD163的表达,以及炎症因子IL-1β、IL-6、IL-12、TGF-α、TGF-β和IL-10的表达,同时进一步检测巨噬细胞极化相关miRNA分子miRNA9、miRNA155和miRNA34a的表达。结果奥氮平能抑制THP-1单独组和共培养组中THP-1的增殖活性(P<0.05);在佛波酯诱导THP-1向巨噬细胞分化的研究中,奥氮平能够显著提高THP-1单独组CD68分子的表达(P<0.05),而在共培养组中CD68、CD80、CD163的表达则均显著升高(P<0.05)。在THP-1单独组中,奥氮平仅能够促进IL-12的表达;在共培养组中,奥氮平使CCL2、CXCL10、IL-1β、IL-10、IL-6、IL-12、TNF-α表达水平显著升高(P<0.05)。在巨噬细胞分化相关miRNA的研究中,在奥氮平作用下,THP-1单独组中miR155和Let7c的表达显著升高(P<0.05),miR146、miR9和miR34a的表达变化无差异;在与MDA-MB-231共培养条件下,miR34a和Let7c显著升高(P<0.05)。结论在肿瘤细胞存在的情况下,奥氮平可参与肿瘤相关巨噬细胞表面分子、炎症因子以及巨噬细胞极化相关miRNA分子表达的调节,这为进一步利用奥氮平治疗乳腺癌提供了新思路。

The regulatory pathways leading to stem-like cells underlie prostate cancer progression

Prostate cancer (PCa) is the most common cause of malignancy in males and the third leading cause of cancer mortality in the United States. The standard care for primary PCa with local invasive disease mainly is surgery and radiation. For patients with distant metastases, androgen deprivation therapy (ADT) is a gold standard. Regardless of a favorable outcome of ADT, patients inevitably relapse to an end-stage castration-resistant prostate cancer (CRPC) leading to mortality. Therefore, revealing the mechanism and identifying cellular components driving aggressive PCa is critical for prognosis and therapeutic intervention. Cancer stem cell (CSC) phenotypes characterized as poor differentiation, cancer initiation with self-renewal capabilities, and therapeutic resistance are proposed to contribute to the onset of CRPC. In this review, we discuss the role of CSC in CRPC with the evidence of CSC phenotypes and the possible underlying mechanisms.

乳腺癌标志物影响卵丘细胞基因表达的生物信息学分析

为探讨乳腺癌患者大概率患有不孕症的情况,本研究应用生物信息学方法来研究乳腺癌标志物对卵丘细胞基因表达的影响。从NCBI的GEO数据库下载有关乳腺癌标志物的基因芯片数据库和卵丘细胞基因表达的基因芯片数据库。利用在线GEO2R筛选出2个数据库的差异基因,在EXCEL表格中进行筛选,对2个结果作韦恩图获得2个数据库差异基因的交集,利用DAVID6.8数据库对差异基因进行GO功能分析和KEGG通路分析,运用String-db数据库构建蛋白之间的相互作用图(PPI),导入Cytoscape3.6.1软件获取关键靶基因。本文通过生物信息学方法从不同角度揭示有关乳腺癌标志物对卵丘细胞差异基因表达的影响,为人类乳腺癌患者生育能力的提高带来新思路。

TIGIT、CD155在乳腺癌组织中的表达及临床意义

目的 观察T细胞免疫球蛋白和免疫受体酪氨酸抑制基序结构域(TIGIT)、白细胞分化抗原155(CD155)在乳腺癌组织中的表达,并探讨其与乳腺癌患者预后的关系。方法 本研究为前瞻性研究,纳入2020年1月至2021年12月三门峡市中心医院收治的乳腺癌患者作为研究对象。所有患者均接受改良根治术治疗,术后检测癌组织及癌旁组织中TIGIT、CD155表达情况,随访1 a观察预后情况。采用Cox生存回归分析TIGIT、CD155表达与乳腺癌患者预后的关系。结果 63例患者失访1例,最终纳入62例。62例患者癌组织中TIGIT阳性32例,CD155阳性43例,TIGIT阳性表达率、CD155阳性表达率均高于癌旁组织(P<0.05)。62例患者术后1 a内发生预后不良16例,占25.81%,预后不良患者癌组织中TIGIT、CD155阳性表达率、TNM分期Ⅲ期占比高于预后良好患者(P<0.05)。Cox回归分析结果显示,TNM分期为Ⅲ期、TIGIT为阳性、CD155为阳性是乳腺癌患者预后不良的危险因子(HR>1,P<0.05)。结论 TIGIT、CD155在乳腺癌患者癌组织中阳性表达率较高,会增加患者预后不良风险。

非小细胞肺癌神经内分泌分化的免疫组化诊断

目的： 探讨非小细胞肺癌神经内分泌分化的免疫组化诊断。方法： 选择三种神经内分泌共同标记ＣＧＡ、ＳＹＮ、ＮＣＮＭ， 应用免疫组化ＬＳＡＢ 法， 观察它们在ＮＳＣＬＣ 中的表达。结果： 在１６３ 例ＮＳＣＬＣ 中， １８ 例（１１－０４ ％ ） 表达ＣＧＡ，６３ 例（３８－６５ ％ ） 表达ＳＹＮ，４０ 例（２４－５４ ％ ） 表达ＮＣＡＭ 。７７ 例（４７－２４ ％ ） ＮＳＣＬＣ 至少有１ 种ＮＥ 标记表达，其中４０ 例只表达１ 种标记，３０ 例表达２ 种标记，仅７ 例３ 种标记均表达。３ 种标记表达彼此相关（ Ｐ＜０－００５） ，但存在差异（ Ｐ＜ ０－００５） 。与腺癌相比，腺鳞癌、鳞癌、大细胞癌染色阳性率较低。结论：选择３ 种ＮＥ 标记可以检测出ＮＳＣＬＣ 的ＮＥ 分化， 但不能以某一种标记的表达与否作为ＮＳＣＬＣ 的ＮＥ 分化诊断， 应联合检测以至少２ 种标记表达定义为ＮＥ 分化，在不同类型的ＮＳＣＬＣ 中腺癌更易出现ＮＥ 分化。

肺鳞状细胞癌伴神经内分泌分化与生物学特性及预后关系探讨

目的:初步探讨肺鳞状细胞癌的神经内分泌分化(neuroendoerine-like differentiation,ND)情况与生物学行为和预后的关系。方法:收集手术后经病理证实为肺原发鳞癌患者120例瘤组织石蜡标本,术前均未接受放化疗。采用免疫组化二步法检测神经元特异性烯醇化酶(NSE),突触素(SYN)和嗜铬素A(CGA)的表达,采用x2检验分析ND与肺鳞癌生物学行为之间的关系,分别采用kaplan-meier生存曲线和COX风险比例模型进行单因素和多因素生存分析。结果:120例肺鳞癌中,CGA阳性率46.7%,NSE阳性率60.8%,SYN阳性率36.7%。鳞癌伴ND阳性率为40.8%。相关性分析结果示3种标志物之间无明显相关性。肺鳞状细胞癌伴ND与肿瘤细胞分化程度及瘤体分期(T分期)有关,肿瘤细胞分化越差,瘤体分期越高,ND阳性率越高。SYN的阳性表达与患者的年龄、性别及肿瘤细胞的分化程度有关,年龄越高,女性,肿瘤细胞分化越差,SYN阳性表达率越高。单因素生存分析结果经Log-Rank检验示SYN(x^2=3.961,P=0.047)及术后化疗(x^2=4.447,P=0.035)与患者的术后生存率有关,而ND与患者术后生存率无明显相关性(x^2=0.524,P=0.469)。Cox多因素生存分析结果示患者的年龄(OR=0.459,P=0.029),SYN阳性表达(OR=2.114,P=0.036),CGA阳性表达(OR=0.558,P=0.066)和术后化疗(OR=1.866,P=0.091)与患者预后相关,可作为可切除肺鳞癌患者的独立预后因素。结论:肺鳞癌伴神经内分泌分化与肿瘤细胞分化程度及瘤体分期有关,SYN和术后化疗是评价可手术患者预后的两个重要指标。

胆固醇抑制剂洛伐他汀对MCF-7细胞增殖分化功能及形态结构的影响

目的 探讨洛伐他汀 (Lovastatin ,LOV)对MCF 7细胞增殖、分化功能及细胞形态、结构的影响。方法 LOV处理细胞 1～ 3d后 ,MTT比色法检测细胞增殖功能 ,流式细胞仪分析细胞周期相分布及细胞凋亡率 ,同时观察细胞形态、结构的改变。结果 LOV可导致细胞形态、结构改变 ;细胞增殖抑制 ,生长阻滞于G0 /G1期 ;同时LOV有诱导该细胞向正常细胞分化的趋势 ,但诱导细胞凋亡的作用不明显。结论 LOV可导致MCF 7细胞形态、功能改变 ,细胞增殖受抑、生长阻滞于G0 /G1期 。

CgA和SYN在非小细胞肺癌中的表达及与预后的关系

目的研究神经内分泌分化（ND）标志物嗜铬蛋白A（CgA）和突触素（SYN）在可切除的非小细胞肺癌（NSCLC）组织中的表达及其与预后的关系。方法2000年1月～2003年1月收治NSCLC且接受手术治疗的患者共123例。收集手术切除标本以及患者临床和术后随访资料等。应用免疫组化二步法检测肿瘤组织标本中CgA和SYN的表达；分别采用Kaplan．Meier生存曲线和Cox风险比例模型进行单因素和多因素生存分析。结果切除的肿瘤组织标本中CgA和SYN的阳性表达率分别为22％和17．9％；SYN阳性表达与肿瘤分化程度相关（P=0．001）；NSCLC伴ND（NSCLC—ND）（CgA和SYN任意一项表达阳性）与患者性别、年龄、吸烟指数、TNM分期和病理类型无明显相关。耐受化疗次数在CgA、SYN呈阳性表达与阴性表达的NSCLC患者之间无明显差异（P=0．406）。Kaplan—Meier生存分析显示，CgA和SYN表达与患者生存期相关；Cox多因素分析表明，肿瘤组织中SYN阳性表达（P=0．001）、TNM分期（P=0．02）和最大直径（P=0．049）是影响患者预后的独立危险因素。结论NSCLC—ND患者预后较差，其标记物SYN可作为临床判断预后的指标之一。

Genistein对乳腺癌细胞致成骨细胞生物学功能改变的影响

目的探讨Genistein对乳腺癌细胞致成骨细胞（osteoblast,OB）增殖、分化和矿化功能的影响,观察Genistein在乳腺癌骨转移的病理条件下是否能调节OB生物学功能。方法源于大鼠颅盖骨的原代OB与50%来自人源性乳腺癌细胞系MDA-MB-231或MCF-7的条件培养基（conditioned medium,CM）共同培养,并加入5×10-7mol/L（G7）、5×10-8mol/L（G8）或5×10-9mol/L（G9）的Genistein进行干预。MTT法观察其对OB增殖的影响;PNPP偶氮法观察其对OB碱性磷酸酶（alkaline phosphatase,ALP）活性的影响;茜素红S（ARS）进行矿化结节染色并计算面积以观察其对OB矿化能力的影响。结果MDA-MB-231和MCF-7细胞条件培养基可显著抑制OB的增殖。用Genistein干预1 d、3 d和5 d后,OB增值率可有不同程度的提高,差异有统计学意义（P〈0.05）。此外,乳腺癌细胞条件培养基可明显下调OB的ALP活性,而用不同浓度的Genistein干预后,OB的ALP活性分别较MDA-MB-231和MCF-7细胞条件培养基组增加22.7%、32.4%、63.5%和27.7%、32.0%、58.3%（P〈0.05）。Genistein还可改善乳腺癌细胞条件培养基对OB矿化能力的抑制,增加OB形成的矿化结节面积。结论在乳腺癌骨转移的病理条件下,Genistein可促进OB的增殖、分化和矿化能力,改善乳腺癌细胞对OB生物学功能的抑制作用。

磁共振联合血清GDF3、CYFRA211、CEA检测在乳腺癌诊断中的临床意义

目的探讨核磁共振联合血清生长分化因子(GDF3)、细胞白蛋白19片段(CYFRA211)、癌胚抗原(CEA)在乳腺癌诊断中的价值。方法选取63例乳腺病变患者作为研究对象,患者均采用核磁共振(MRI)检测,血清GDF3、CYFRA211、CEA测定采用酶联免疫法,观察患者手术病理诊断结果。以手术病理诊断结果作为参考,对比MRI诊断、血清诊断以及二者联合诊断效能以及受试者工作特征(ROC)曲线下面积(AUC)。结果 MRI检测乳腺癌患者诊断符合率91.18%,灵敏度91.30%,特异度90.91%,误诊率8.33%,漏诊率8.70%。血清GDF3、CYFRA211、CEA诊断符合率85.29%,灵敏度84.78%,特异度86.36%,误诊率15.22%,漏诊率13.64%。联合检测诊断符合率97.06%,灵敏度95.65%,特异度100.00%,漏诊率4.35%。联合诊断符合率、敏感度以及AUC值均高于单独检测,差异有统计学意义(P<0.05),而特异度无显著差异(P>0.05)。结论乳腺癌患者采用MRI联合血清GDF3、CYFRA211、CEA检测的诊断效果较好,对于乳腺癌的临床诊断具有一定的参考价值。

乳腺癌术前中医辨证与肿瘤增殖因子相关性研究

目的:探讨乳腺癌术前中医辨证与肿瘤增殖因子DNA倍体、S期细胞比例及Ki-67表达的相关性。方法:对160例乳腺癌患者术前中医辨证分型,术后手术标本进行DNA倍体、S期细胞比例、Ki-67检测,并做相关性分析。结果:乳腺癌患者术前辨证为正虚毒炽者DNA倍体异倍体、S期细胞比例为高度及Ki-67阳性出现率高于肝郁痰凝组和冲任失调组。经统计学处理,有显著差异(P<0.05)。结论:乳腺癌术前中医辨证与肿瘤增殖因子DNA倍体、S期细胞比例及Ki-67表达有相关性,随着患者中医辨证为肝郁痰凝证、冲任失调证、正虚毒炽证的不同,肿瘤的增殖能力依次增强,预后也更差。

乳腺癌干细胞的富集及相关基因表达

目的通过微球体培养富集乳腺癌干细胞,并检测其相关基因的表达。方法将MCF-7乳腺癌细胞进行第一代微球体培养,7d后消化微球体获得单细胞悬液,取部分细胞进行第二代微球体培养,另一部分细胞则在胶原底物进行分化培养。于培养第11天收集第二代微球体细胞、分化细胞,采用流式细胞术进行干细胞比例分析;同时应用Real-time PCR技术检测β-catenin、CXCR4、SOX2、ALDH3A1 mRNA表达。结果流式细胞分析显示,微球体细胞和分化细胞的侧亚群细胞比例分别为(7.36±0.54)%和(4.32±0.42)%(P<0.05),CD55high比例分别为(17.41±1.09)%和(4.47±0.33)%(P<0.05)。Real-time PCR检测结果表明,微球体细胞与分化细胞相比,β-catenin、CXCR4、SOX2及ALDH3A1 mRNA表达分别上调了约1.5、5.0、4.0和5.7倍(P<0.01)。结论微球体细胞富集了较高比例的乳腺癌干细胞,且β-catenin、CXCR4、SOX2、ALDH3A1 mRNA呈现高表达。

乳腺癌高表达基因1对前列腺癌细胞神经内分泌转化的影响

目的观察乳腺癌高表达基因1(BCOX1)在前列腺癌细胞神经内分泌转化中的表达水平和生物功能。方法构建去雄激素诱导的神经内分泌样前列腺癌细胞模型。并检测BCOX1在LNCaP和AI-NEPC细胞中的表达差异。RNA干扰技术下调BCOX1在AI-NEPC细胞中表达后,检测神经内分泌细胞标记物NSE和CHGA的表达,并检测STAT3的表达和磷酸化水平。结果在AI-NEPC中BCOX1蛋白表达水平与LNCaP细胞相比明显增加。BCOX1mRNA水平在AI-NEPC中与LNCaP细胞相比显著增高(P<0.05)。shRNA转染组中NSE和CHGA表达水平与未转染组和空载体转染组比较均明显下降(P<0.05)。shRNA转染组中STAT3和磷酸化STAT3的表达水平较空载体转染组明显下调(P<0.05)。结论 BCOX1在神经内分泌分化的前列腺癌细胞中高表达,下调BCOX1的表达可以影响前列腺癌细胞的神经内分泌样表型,并抑制STAT3的表达和磷酸化。

乳宁Ⅱ号方对小鼠Ca761乳腺癌移植瘤细胞周期及p53和ras表达的影响

目的:通过研究乳宁Ⅱ号方对小鼠Ca761乳腺癌移植瘤细胞周期及p53、ras表达的影响,探讨乳宁Ⅱ号方抑制乳腺癌生长的机制。方法:建立小鼠Ca761 乳腺癌移植模型,小鼠造模后随机分成生理盐水对照组、环磷酰胺(cyclophosphamide, CTX)治疗组、乳宁Ⅱ号方治疗组及乳宁Ⅱ号方加CTX 治疗组,每组12只。用流式细胞术检测经不同方法处理的荷瘤小鼠移植瘤细胞的细胞周期,同时运用免疫组化法测定移植瘤组织中p53和ras癌基因蛋白的表达,并进行组间比较。结果:乳宁Ⅱ号方治疗组、CTX治疗组与乳宁Ⅱ号方加CTX治疗组S期肿瘤细胞与生理盐水对照组比较明显减少(P<0.05);乳宁Ⅱ号方治疗组G0 G1期肿瘤细胞百分比低于CTX治疗组(P<0.05),而G2 M期细胞百分比高于CTX治疗组(P<0.05);乳宁Ⅱ号方治疗组与乳宁Ⅱ号方加CTX治疗组p53的表达均显著低于生理盐水对照组(P<0.05),而CTX对p53表达的影响不显著(P>0.05);乳宁Ⅱ号方治疗组、CTX治疗组与乳宁Ⅱ号方加CTX治疗组ras的表达均低于生理盐水对照组(P<0.05)。结论:乳宁Ⅱ号方可通过调控小鼠Ca761 乳腺癌移植瘤细胞的细胞周期来影响肿瘤的生长,这一作用与其调控肿瘤组织p53和ras基因表达有关。

CD44^+CD24^(-/low)乳腺癌干细胞的流式分选及其肿瘤干细胞特性的初步鉴定

目的:通过表面标志分选法富集乳腺癌干细胞,并初步鉴定其肿瘤干细胞特性。方法:采用流式细胞分选术从人乳腺癌细胞系MCF-7中分选CD44+CD24-/low乳腺癌干细胞,并进行干细胞比例分析;用免疫荧光法检测、比较分选获得的细胞和对照细胞的干性和分化标记物Oct-4、SOX-2、CK-18和α-SMA的表达状态。结果:分选获得的CD44+CD24-/low乳腺癌干细胞阳性比例达90%以上;免疫荧光检测结果显示,CD44+CD24-/low细胞亚群比non-CD44+CD24-/low细胞亚群高表达干细胞转录因子Oct-4、SOX-2,低表达分化因子CK-18、α-SMA;体外实验表明,CD44+CD24-/low细胞亚群具有更强的成球生长能力,并具有双向分化潜能。结论:CD44+CD24-/low表面标记物分选的方法可以富集高纯度的乳腺癌干细胞,且呈现干性因子Oct-4和SOX-2高表达。

TGF-α诱导前列腺癌DU145细胞神经内分泌分化和增强DU145对顺铂的化疗耐药性

目的:研究TGF-α是否具有EGF相似的可诱导前列腺癌DU145细胞发生神经内分泌分化(neuroendocrine differentiation,NED)的作用,并探讨TGF-α诱导的前列腺癌DU145细胞神经内分泌分化对肿瘤化疗耐药性的影响。方法:将接受不同培养液处理的DU145细胞分为3组:2%FBS组、2%FBS+TGF-α5 ng/mL组和2%FBS+TGF-α10 ng/mL组。显微镜下观察TGF-α处理后DU145细胞的形态变化;Real time RT-PCR法检测神经特异性烯醇化酶(neuron-specific enolase,NSE)mRNA的表达水平;Western印迹检测NSE,P-糖蛋白(P-glycoprotein,P-gp),多药耐药相关蛋白1(multiple drug resistance protein,MRP1)和Bcl-2蛋白的表达水平。流式细胞术分析TGF-α(5μg/mL)处理后DU145细胞周期的变化;MTT比色法测定TGF-α(5μg/mL)对DU145细胞顺铂化疗耐药性的影响。结果:同2%FBS组相比,2%FBS+TGF-α处理组的DU145细胞出现多形性,细胞伪足伸出;NED标志物NSEmRNA的表达水平升高,分别为上调了(3.6±0.5)倍(P<0.05)和(10.1±0.1)倍(P<0.01);细胞的NSE,Bcl-2和MRP1蛋白的表达水平也明显增加,但未检测到P-gp蛋白的表达;同时,TGF-α(5μg/mL)处理后DU145细胞的G1期细胞比例降低,S期和G2/M期细胞比例升高;顺铂的化疗耐药性增加。结论:TGF-α也可诱导前列腺癌DU145细胞神经内分泌分化增强,从而进一步增强DU145细胞对顺铂的化疗耐药性。