Role of TLR4 for paclitaxel chemotherapy in human epithelial ovarian cancer cells

Background  Paclitaxel has been reported to be a ligand to Toll like receptor 4 (TLR4). Myeloid differentiation factor 88(MyD88) was described as a myeloid differentiation primary response gene. TLR4 signalling owns two pathways: MyD88-dependent and MyD88-independent pathways. XIAP is a key member of the inhibitor of apoptosis protein family. Akt is a major downstream target of growth factor receptor tyrosine kinases, which negatively regulates apoptotic pathways through phosphorylation (pAkt). The aim of the present study is to investigate the role of TLR4 in paclitaxel resistance of ovarian cancer cells.
Materials and methods  We reconstructed the RNA interference expression vector, pGenesil-1-U6 specifically targeting TLR4 mRNA, which was stable transfected into the human ovarian cancer cell line SKOV3 (MyD88-positive expression) and A2780 (MyD88-negative expression). Cell proliferation, cell cycle distribution and cell apoptosis were assessed in the cells transfected with scramble control shRNA (SKOV3/shControl, A2780/shControl) and TLR4 shRNA (SKOV3/shTLR4, A2780/shTLR4) to explore the possible functions of TLR4 in ovarian cancer cells growth. The expression of TLR4, MyD88, XIAP, Akt and pAkt was analysed by Western blot analysis.
Results  A knockdown of TLR4 levels down-regulated the expression of XIAP and pAkt. And it restored the inhibitory effect of paclitaxel on cell proliferation and impeding cell cycle progression in SKOV3 cells.
Conclusions  It suggests that TLR4 negatively regulates paclitaxel chemotherapy and MyD88 is an essential downstream factor to TLR4 signalling for this resistance. Knockdown of TLR4 induces paclitaxel chemosensitivity which might depress the Akt pathway. The TLR4-MyD88 signalling represents an important source to promote tumour growth.     

Severe skin toxicity observed with the combination of capecitabine and weekly paclitaxel in metastatic breast cancer patients

Background  Skin toxicity has recently been recognized as a dose-limiting toxicity of antineoplastic therapy. Discussion  We report severe skin toxicity observed in anthracycline-pretreated metastatic breast cancer patients receiving the combination of capecitabine and weekly paclitaxel.     

表柔比星联合多西他赛或紫杉醇治疗乳腺癌效果差异分析

目的探讨表柔比星联合多西他赛或紫杉醇治疗乳腺癌的效果差异。方法选取2018年2月至2019年7月本院收治的86例乳腺癌患者,根据动态随机分组法将其分为对照组(n=43)和观察组(n=43),对照组采用表柔比星联合紫杉醇治疗,观察组采用表柔比星联合多西他赛治疗,比较两组临床缓解率、1年内复发率、生存时间以及不良反应发生率。结果两组患者临床缓解率差异经统计学软件检验显示无统计学意义(P>0.05);观察组1年内复发率和不良反应发生率低于对照组,生存时间长于对照组,差异经统计学软件检验显示有统计学意义(P<0.05)。结论表柔比星联合多西他赛或紫杉醇对乳腺癌的治疗效果相近,但联合多西他赛的不良反应风险更低,还可延长患者生存时间。     

靶向TLR4治疗结直肠癌的可行性探讨

慢性炎症与肿瘤的发生发展密切相关.介导炎症的Toll样受体(toll-like receptor, TLR)家族是感受病原体入侵的一类模式识别受体,其中TLR4与结直肠肿瘤发生发展的关系最为明确.本文着重对TLR4与结直肠癌的发病机制、预后以及治疗进行介绍,并对TLR4作为结直肠癌治疗靶标的可行性进行探讨.     

Prospective assessment of chemotherapy-induced peripheral neuropathy due to weekly paclitaxel in patients with advanced or metastatic breast cancer (CSP-HOR 02 study)

Goals of work  The aim of this study was to prospectively evaluate chemotherapy-induced peripheral neuropathy (CIPN) using a patient-based instrument, the Patient Neurotoxicity Questionnaire (PNQ) and a physician-based instrument, the National Cancer Institute-Common Toxicity Criteria (NCI-CTC) in patients with advanced or metastatic breast cancer who were treated with weekly paclitaxel. Materials and methods  CIPN symptoms were prospectively assessed in 35 patients using the PNQ, NCI-CTC, and the Functional Assessment of Cancer Therapy (FACT)-Taxane including neurotoxicity component (Ntx) at the baseline, and 8 and 16 weeks after starting chemotherapy. Results  For sensory neuropathy symptoms, the reported incidence of CIPN was significantly increased during active treatment in terms of both the PNQ and NCI-CTC assessments. In contrast, there was a notable increase of patient motor neuropathy symptoms that were elucidated only by the PNQ. The PNQ grades of CIPN were widely distributed in the patient population as compared with the NCI-CTC grades for both sensory and motor neuropathy. The sensory PNQ grade was correlated with sensory NCI-CTC grade (r = 0.58) and Ntx (r = 0.51), and the motor PNQ grade was correlated with Ntx (r = 0.57). Conclusions  The PNQ appears to be more sensitive and responsive than the NCI-CTC for CIPN; the PNQ appears to have diagnostic validity for evaluating CIPN in patients who are receiving neurotoxic chemotherapy.     

Inhibition of autophagy sensitizes MDR-phenotype ovarian cancer SKVCR cells to chemotherapy

Autophagyis an intracellular lysosomal degradation pathway where its primary function is to allow cells to survive under stressful conditions. Autophagy is, however, a double-edge sword that can either promote cell survival or cell death.Chemoresistanceis a major challenge in the clinical treatment of ovarian cancer, of which the underlying mechanisms remain unknown.ObjectiveThe aim of the present study was to explore the role of autophagy in vincristine (VCR) resistant ovarian cancer cells.MethodsThe SKOV3 parental cell line and SKVCR, the VCR-resistant ovarian carcinoma cells were used. 3-MA (3-Methyladenine) and CQ (Chloroquine) were also used as autophagy inhibitors. CCK8 (Cell Counting Kit-8) was used to detect cell viability, quantitative real-time PCR and Western blot were used to detect the expressions of mRNA and protein, MDC staining and flow cytometry were used to detect autophagy and apoptosis, respectively.ResultsCompared with parental SKOV3 cells, SKVCR cells showed Multidrug Resistance (MDR). SKVCR cells demonstrated higher autophagy levels than SKOV3 cells, which could be inhibited by 3-MA and CQ. In SKVCR cells, VCR increased apoptosis levels further, 3-MA and CQ inhibited autophagy and potentiated the cytotoxicity by VCR. Moreover, 3-MA and CQ overcame the acquired VCR resistance in SKVCR cells by enhancing VCR-induced cytotoxicity, and promote apoptosis.ConclusionsOur data indicate that autophagy has a protective role in the multi-drug resistant SKVCR cells. The inhibition of autophagy increases the killing effects of VCR by increasing apoptosis and inhibiting autophagy, suggesting a better strategy for the treatment of drug-resistant SKVCR cells.     

Mechanism of TLR4 mediated immune effect in transfusion-induced acute lung injury based on Slit2/Robo4 signaling pathway

BackgroundTransfusion-related acute lung injury (TRALI) is the infusion of blood or blood system.ObjectiveTo explore the mechanism of TLR4-mediated T cell immune effect in TRALI.MethodsIn this animal study, a mouse model of LPS-induced TRALI was established. Sixty adult C57/BL6 mice (wild-type, WT) were randomly divided into 5 groups: 1) normal WT type, 2) LPS control group of WT type lipopolysaccharide, 3) WT type TRALI group (LPS + MHC-I mAb), 4) (TLR4 antibody) lipopolysaccharide LPS control group, 5) (TLR4 antibody) TRALI group (LPS + MHC-I mAb). Mice were injected with LPS (0.1 mg/kg) and MHC-I mAb (2 mg/kg) into the tail vein. H&E staining was performed to detect pathological features. The myeloperoxidase (MPO) activity and the level of inflammatory cytokines in lung tissue homogenate supernatant were measured. Blood, spleen single-cell suspension, and bronchoalveolar lavage fluid were collected to detect the ratio of Treg and Th17 cells by flow cytometry. RT-PCR and WB were used to detect mRNA or protein expression.ResultsTLR4 mAb treatment alleviated the pathogenesis of LPS-induced TRALI in vivo, the MPO activity, and the level of proinflammatory factors in lung tissues. TLR4 exerted its function by changing of Treg/Th17 ratio via the SLIT2/ROBO4 signaling pathway and downregulating CDH5 and SETSIP.ConclusionTLR4 mediates immune response in the LPS-induced TRALI model through the SLIT2/ROBO4 signaling pathway.     

Notch1信号通路在乳腺癌中的研究进展

乳腺癌是女性最常见、死亡率最高的恶性肿瘤之一,近年来其发病率不断升高。研究发现Notch1信号通路在乳腺癌中起着重要的作用。研究发现Notch1在正常乳腺组织和乳腺良性病变以及乳腺癌中均有表达,在乳腺浸润性导管癌中的表达率明显高于导管原位癌,其表达与乳腺癌HER-2亚型有关,在HER-2阳性型中的Notch1的阳性表达率明显高于其他亚型。研究认为Notch1阳性表达率高与其乳腺癌的分化程度低、分期晚、淋巴结转移阳性率高以及脑转移有关,预示着乳腺癌的预后差,并被认为可作为判断预后的临床指标。此外研究发现Notch1的高表达可导致乳腺癌对阿霉素与紫杉醇的耐药。研究发现Notch1高表达与乳腺癌侵袭强、凋亡抑制有关。研究还发现Notch1的表达影响乳腺癌干细胞的数量,是维持乳腺癌干细胞恶性表型的重要因子。且Notch1与VEGF、c-myc、CCL2、TRB3、MMP2等相关基因表达及肿瘤微环境相关。针对Notch1的靶向治疗的研究有Notch1单克隆抗体、三氧化二砷以及金雀异黄素等药物,以及Notch1相关通路的靶向治疗药物,均可抑制乳腺癌细胞的生长,对乳腺癌有抗肿瘤作用。     

库普弗细胞对急性肝衰竭模型鼠TLR4表达的影响

目的:探讨抑制库普弗细胞对急性肝衰竭模型鼠Toll样受体4(Toll-like receptor 4.TLR4)表达的影响.方法:80只SD雄性大鼠随机分为正常对照组、氯化钆(gadolinium chloride,GdCl3)组、内毒素(LPS)组、GdCl3+LPS组.免疫组化检测EDl表达,观察GdCl3对库普弗细胞的抑制程度,并检测血清ALT、TNFα、IL-1β的水平,Western blot分析肝组织TLR4的表达.结果:GdCl3组肝组织中EDl免疫反应阳性细胞与正常对照组相比明显减少(P＜0.05),LPS组EDl阳性细胞数量最多,GdCl3+LPS组EDl阳性细胞数少于LPS组(P＜0.05).GdCl3+LPS组AIJT、TNFct、IL-1β水平高于正常对照组,但低于LPS组(P＜0.05).GdCl3+LPS组大鼠肝脏TLR4表达与LPS组相比较弱(P＜0.05),与正常对照组相比差异无统计学意义(P＞0.05).结论:抑制库普弗细胞激活可减少肝组织11LR4的表达,库普弗细胞在急性肝衰竭中发挥重要作用.     

肿瘤相关巨噬细胞诱导乳腺癌耐药的实验研究

目的探索肿瘤相关巨噬细胞(TAM)在乳腺肿瘤细胞耐药中的作用及其耐药机制。方法通过佛波酯(PMA)、白细胞介素4(IL-4)、白细胞介素13(IL-13)刺激人单核细胞白血病细胞系THP-1,建立TAM模型在体外与乳腺癌细胞共培养,运用流式细胞术(FCM)检测TAM表面分子CD14、CD204;使用噻唑蓝(MTT)比色法检测紫杉醇对乳腺癌细胞系T47D、BT-549的有效杀伤浓度和时间,以及乳腺癌细胞单独培养、乳腺癌细胞与TAM/TAM上清共培养下乳腺癌细胞的杀伤;运用Western blot检测肿瘤细胞的磷酸化信号转导与转录激活因子3(p-STAT3)、磷酸化c-jun氨基末端激酶(p-JNK)信号通路变化。结果 THP-1细胞经PMA、IL-4、IL-13诱导分化为TAM,其细胞表面表达CD14、CD204(表达率分别为31.52%±9.39%、48.21%±8.76%)。TAM/TAM上清与肿瘤细胞共培养均可显著削弱紫杉醇对肿瘤细胞的杀伤(T47D相对存活率由43.52%±9.40%提高至92.68%±2.32%/92.20%±2.10%,BT-549相对存活率由63.08%±2.71%提高至77.96%±2.64%/79.55%±2.35%,P0.05),明显下调其凋亡信号p-JNK(P0.05),同时显著上调p-STAT3(P0.05)。结论成功建立TAM模型,TAM介导乳腺肿瘤细胞耐药,其机制可能与TAM分泌的细胞因子影响肿瘤细胞STAT3、JNK信号分子磷酸化有关。     

Cardiac toxicity assessment in locally advanced breast cancer treated neoadjuvantly with doxorubicin/paclitaxel regimen

Background The psychological difficulty of accepting a mastectomy for locally advanced breast cancer (LABC) justifies the use of chemotherapy as neoadjuvant primary treatment. The aim of this prospective study was to assess the efficacy of the doxorubicin/paclitaxel (AT) schedule neoadjuvantly administered in terms of response rates and survival in patients with LABC, with a special focus on cardiac toxicity.Patients and method All patients were treated by doxorubicin (60 mg/m² i.v.) bolus followed by paclitaxel (200 mg/m²) as a 3-h infusion. Treatment was repeated every 3 weeks for four or six courses and followed by surgery, radiotherapy, and hormonotherapy for patients with positive hormonal receptors. Patients with significant cardiovascular history or ECG abnormalities were not eligible for the study. Measurements of left ventricular ejection fraction (LVEF) were performed at baseline and at the end of chemotherapy.Results From 1998 to 2001, 34 consecutive patients followed up in our institution were entered into this study. Median age was 49 years (range, 32–68 years). Seventeen patients had stage IIB, 5 patients stage IIIA, and 12 patients stage IIIB disease. Twenty-one patients underwent conservative surgery, 7 radical surgery, and 6 patients no surgery due to metastatic disease occurring during treatment. An objective clinical response was noted in 22 (65%) of 34 patients (6 patients with histological complete response, 10 patients with rare malignant cells, and 6 patients with a partial response), 6 patients presented a progressive disease, and 8 patients a stable disease. Twenty-four patients have kept normal cardiac function, 7 patients had a cardiac toxicity as defined by the institution [4 (24%) of 17 patients received 360 mg/m² of doxorubicin (A), 2 of 4 presented congestive heart failure (CHF), and 3 (21%) of 14 patients received 240 mg/m² of A without CHF]. Three patients did not receive four or six cycles as initially planned due to the progressive disease during the chemotherapy courses. These patients were excluded from the final analysis, particularly cardiac toxicity analysis. At time of median follow-up (42 months), 28 of 34 patients were alive (one death due to CHF, five others due to progressive disease).Conclusion The AT regimen in neoadjuvant treatment for LABC remains efficient, but cardiac toxicity reported in this study underlies the necessity to optimize the schedule of AT combination.     

临床护理路径在乳腺癌围手术期护理中应用的效果评价

目的 探讨临床护理路径在乳腺癌围手术期护理中的应用效果,特别是在乳腺癌术后及出院后患肢功能锻炼中的指导作用.方法 严格按照临床护理路径的准入和退出标准,选取普外科乳腺癌患者102例,按照入院单双日随机分为实验组50例和对照组52例.实验组采取临床护理路径指导下的"一线式"护理服务模式,对照组采用传统的整体护理模式.统计2组患者对疾病相关知识的掌握程度、出院后自护能力及患者对护理服务满意度,并进行统计分析.结果 实验组患者对围手术期相关知识的掌握程度和出院后自护能力评分明显高于对照组,且患者对护理服务满意度达98.92%,显著高于对照组.结论 临床护理路径指导下的"一线式"护理服务模式在乳腺癌患者围手术期护理中的应用,取得了良好的效果,督促了乳腺癌患者术后的患肢功能锻炼,促进了疾病的恢复,缩短了住院时间,节省了住院费用.

Abstract：

Objective To explore the effect of clinical nursing pathway (CNP) in perioperative nursing for breast cancer,especially the instructive important role in the rehabilitative excises postoperation and after discharge.Methods In accordance with the standard for the entry and exit of the CNP,102 patients with breast cancer were randomly divided into the experimental group (50 patients) and the control group (52 patients),the experimental group received the care of clinical nursing pathway and the control group received the traditional care.The effect of clinical nursing pathway on the following aspects such as the level of mastering knowledge about hospitalization,self-care ability after discharge,the degree of satisfaction about nursing were compared between the two groups.Results Compared with the control group,the level of mastering knowledge about hospitalization and self-care ability after discharge in the experimental group were significantly better,the degree of satisfaction about nursing reached 98.92%,which was higher than that of the experimental group.Conclusions Application of the full care model with CNP demonstrated good effect on the perioperative nursing for breast cancer,it encourages patients to do the limb functional training,promotes the recovery of disease,and then shortens hospital stay and saves hospital costs.     

HE4、CAl53联合检测在乳腺癌辅助诊断中的应用

目的研究血清人附睾上皮分泌蛋白4（HE4）、癌抗原153（CAl53）联合检测在乳腺癌辅助诊断中的价值。方法收集46例乳腺癌患者、52例乳腺良性疾病患者以及50名正常对照者血清,采用酶联免疫吸附试验（ELISA）检测血清HE4含量、电化学发光法检测CAl53含量。采用受试者工作特征（ROC）曲线确定HE4和CA153的临界值。分别计算HE4和CA153单项检测和联合检测的敏感性和准确性并作比较。结果乳腺癌组HE4和CAl53水平分别是49.0（41.7～67.4）pmol/L和20.6（13.4～28.7）U/mL,均明显高于乳腺良性疾病组及正常对照组（P〈0.01）。经ROC曲线确定HE4和CA153的临界值分别为62.5 pmol/L和21.6 U/mL,乳腺癌HE4、CAl53联合检测的敏感性为63.0%,准确性为82.4%,均高于单项检测。结论联合检测CAl53和HE4可以明显提高乳腺癌诊断的准确性及敏感性。     

TMPRSS4通过上皮间质转化促进乳腺癌细胞的增殖、侵袭和转移

目的通过检测Ⅱ型跨膜丝氨酸蛋白酶4（TMPRSS4）在乳腺癌细胞中的表达情况,分析其在乳腺癌细胞增殖、侵袭和转移过程中的作用及其与上皮间质转化（EMT）的关系。 方法采用实时荧光定量PCR（qRT-PCR）和Western blot法检测5种不同乳腺癌细胞系中TMPRSS4 mRNA和蛋白水平的表达情况。将过表达质粒转染至乳腺癌细胞中,采用qRT-PCR方法检测转染效率,并通过MTS和EdU细胞增殖实验、Transwell和Matrigel细胞迁移和侵袭实验,研究过表达TMPRSS4对乳腺癌细胞增殖、侵袭和迁移的作用。采用qRT-PCR和Western blot法检测过表达TMPRSS4后细胞EMT相关基因E-cadherin、Vimentin、Claudin-1、Slug、ZEB1的表达变化。 结果TMPRSS4在乳腺癌MDA-MB-468和MDA-MB-231细胞系中高表达。MTS实验显示TMPRSS4过表达组乳腺癌细胞MDA-MB-468和MDA-MB-231的增殖能力与阴性对照组相比明显增加,差异具有统计学意义（P=0.039和0.038）,EdU实验进一步证实过表达TMPRSS4促进乳腺癌细胞的增殖,MDA-MB-468细胞和MDA-MB-231细胞的增殖率与阴性对照组比较差异具有统计学意义（P=0.001和0.008）。Transwell实验显示,TMPRSS4过表达组MDA-MB-468细胞和MDA-MB-231细胞的迁移能力与阴性对照组比较明显提高,差异具有统计学意义（p均=0.001）。Matrigel实验显示,TMPRSS4过表达组MDA-MB-468细胞和MDA-MB-231细胞的侵袭浸润能力与阴性对照组比较明显提高,差异具有统计学意义（P=0.012和0.000）。与阴性对照组比较,过表达TMPRSS4后,EMT相关基因包括上皮性标记E-cadherin和Claudin-1的表达均显著下降,而间质标记Vimentin和Slug的表达均明显提高,差异具有统计学意义（P分别=0.024,0.003,0.002和0.012）。 结论TMPRSS4参与了乳腺癌EMT的发生过程,并通过EMT促进乳腺癌细胞的增殖、侵袭和转移。     

Paeoniflorin inhibits proliferation and invasion of breast cancer cells through suppressing Notch-1 signaling pathway

Paeoniflorin (PF), one of the major active ingredients of Chinese peony, was reported to possess anti-tumor effect. However, the role of PF in breast cancer remains to be clarified. Therefore, in this context, the present study investigated the effects of PF on breast cancer cell proliferation and invasion, as well as the underlying mechanism. Our results found that PF suppressed the proliferation and invasion of breast cancer cells. We further demonstrated that PF down-regulated the expression of Notch-1; in addition, overexpression of Notch-1 reversed PF-inhibited proliferation and invasion, and knockdown of Notch-1 enhanced PF-inhibited proliferation and invasion in breast cancer cells. In conclusion, the present study suggests that PF inhibits proliferation and invasion of breast cancer cells through suppressing Notch-1 signaling pathway. Therefore, PF may represent a chemopreventive and/or therapeutic agent in the prevention of breast cancer.     

HE4、CAl53联合检测在乳腺癌辅助诊断中的应用

目的研究血清人附睾上皮分泌蛋白4(HE4)、癌抗原153(CAl53)联合检测在乳腺癌辅助诊断中的价值。方法收集46例乳腺癌患者、52例乳腺良性疾病患者以及50名正常对照者血清,采用酶联免疫吸附试验(ELISA)检测血清HE4含量、电化学发光法检测CAl53含量。采用受试者工作特征(ROC)曲线确定HE4和CA153的临界值。分别计算HE4和CA153单项检测和联合检测的敏感性和准确性并作比较。结果乳腺癌组HE4和CAl53水平分别是49.0(41.7～67.4)pmol/L和20.6(13.4～28.7)U/mL,均明显高于乳腺良性疾病组及正常对照组(P<0.01)。经ROC曲线确定HE4和CA153的临界值分别为62.5 pmol/L和21.6 U/mL,乳腺癌HE4、CAl53联合检测的敏感性为63.0%,准确性为82.4%,均高于单项检测。结论联合检测CAl53和HE4可以明显提高乳腺癌诊断的准确性及敏感性。     

乳腺癌组织趋化因子受体CXCR4的表达与意义

目的探讨趋化因子受体CXCR4在乳腺癌组织中的表达及其临床意义。方法用免疫组织化学方法检测45例乳腺癌及10例乳腺纤维腺瘤组织中趋化因子受体CXCR4的表达,并对原位癌与发生转移的癌组织中的CXCR4的表达率进行比较。结果在乳腺癌组织上检测到趋化因子受体CXCR4的表达（表达率为62．2％）,其中癌组织发生转移的CXCR4的表达率为69．4％,未发生转移的表达率为33．3％,而乳腺纤维腺瘤组织中未发现CXCR4的表达。结论乳腺癌组织上有趋化因子受体CXCR4的表达,其表达可能在乳腺癌的发生、发展和转移中起重要作用。