Liver injury in COVID-19: Holds ferritinophagy-mediated ferroptosis accountable

Even in patients without a history of liver disease, liver injury caused by coronavirus disease 2019 (COVID-19) is gradually becoming more common. However, the precise pathophysiological mechanisms behind COVID-19's liver pathogenicity are still not fully understood. We hypothesize that inflammation may become worse by cytokine storms caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Elevated ferritin levels can initiate ferritinophagy mediated by nuclear receptor coactivator 4 (NCOA4), which leads to iron elevation, and ferroptosis. In COVID-19 patients, ferroptosis can be restricted to reduce disease severity and liver damage by targeting NCOA4-mediated ferritinophagy. To confirm the role of ferritinophagy-mediated ferroptosis in SARS-CoV-2 infection, further research is required.     

Smoothened antagonists reverse taxane resistance in ovarian cancer

The hedgehog pathway has been implicated in the formation and maintenance of a variety of malignancies, including ovarian cancer; however, it is unknown whether hedgehog signaling is involved in ovarian cancer chemoresistance. The goal of this study was to determine the effects of antagonizing the hedgehog receptor, Smoothened (Smo), on chemotherapy response in ovarian cancer. Expression of hedgehog pathway members was assessed in three pairs of parental and chemotherapy-resistant ovarian cancer cell lines (A2780ip2/A2780cp20, SKOV3ip1/SKOV3TRip2, HeyA8/HeyA8MDR) using quantitative PCR and Western blot analysis. Cell lines were exposed to increasing concentrations of two different Smo antagonists (cyclopamine, LDE225) alone and in combination with carboplatin or paclitaxel. Selective knockdown of Smo, Gli1, or Gli2 was achieved using siRNA constructs. Cell viability was assessed by MTT assay. A2780cp20 and SKOV3TRip2 orthotopic xenografts were treated with vehicle, LDE225, paclitaxel, or combination therapy. Chemoresistant cell lines showed higher expression (>2-fold, P < 0.05) of hedgehog signaling components compared with their respective parental lines. Smo antagonists sensitized chemotherapy-resistant cell lines to paclitaxel, but not to carboplatin. LDE225 treatment also increased sensitivity of ALDH-positive cells to paclitaxel. A2780cp20 and SKOV3TRip2 xenografts treated with combined LDE225 and paclitaxel had significantly less tumor burden than those treated with vehicle or either agent alone. Increased taxane sensitivity seems to be mediated by a decrease in P-glycoprotein (MDR1) expression. Selective knockdown of Smo, Gli1, or Gli2 all increased taxane sensitivity. Smo antagonists reverse taxane resistance in chemoresistant ovarian cancer models, suggesting combined anti-hedgehog and chemotherapies could provide a useful therapeutic strategy for ovarian cancer.     

超声联合维拉帕米逆转卵巢癌细胞阿霉素耐受性

目的　探讨超声协同维拉帕米逆转卵巢癌细胞阿霉素耐受性的可能性及其机理。方法　制备卵巢癌细胞阿霉素耐药株 SKOV3/ ADR,并将其分为对照组、超声辐照组 (US组 )、阿霉素组 (ADR组 )、 ADR US组、维拉帕米组 (Vp组 )和 Vp US组 6个组 ,测定耐药细胞株在不同条件下对抗癌药阿霉素的反应性 (计算癌细胞存活率 )及细胞内阿霉素的聚集量。结果　癌细胞对阿霉素的反应性与对照组相比 ,US组细胞存活率无明显降低 (P>0 .0 5 ) ;ADR US组、 Vp组和 Vp US组细胞存活率较 ADR组降低 (P<0 .0 1、 P<0 .0 1、 P<0 .0 1)。 Vp组与 ADR US组差异无显著性 (P>0 .0 5 )。细胞内阿霉素的聚集量 ,ADR US组、 Vp组和 Vp US组与 ADR组相比明显增加 (P<0 .0 5、 P<0 .0 1、 P<0 .0 1) ,Vp US组较 ADR US组和 Vp组增加 (P<0 .0 5、 P<0 .0 5 )。结论　超声可与耐药修饰剂维拉帕米协同作用 ,逆转卵巢癌细胞对阿霉素的耐受性 ,其作用是通过提高细胞内药物聚集量实现的     

沉默缺氧诱导因子-1α对卵巢癌化疗敏感性的影响

目的观察沉默缺氧诱导因子HIF-1α对卵巢癌细胞化疗敏感性的相关影响并探讨其机理。方法构建HIF-1α基因短发夹RNA真核表达质粒,并转染卵巢癌细胞SKOV3（人卵巢浆液性囊腺癌细胞株）。实验可分为空白组,非特异对照组（转染pNonspecific-siRNA,即SKOV3NS）,目的siRNA组（转染pHIF-siRNA,即SKOV3siRNA）3组,用RT-PCR及Western Blot法分别检测各组HIF-1α基因的mRNA和蛋白的表达水平。细胞经过20μmol/L顺铂作用后,用MTT法检测细胞生长抑制率,流式细胞术（FCS）检测细胞凋亡率。结果 SKOV3siRNA转染组HIF-1α基因在mRNA水平和蛋白水平表达均明显降低,SKOV3siRNA组肿瘤细胞的生长抑制率和细胞凋亡率均明显增高（P〈0.05）。结论 RNA干扰技术沉默HIF-1α能够有效地抑制卵巢癌SKOV3细胞HIF-1α基因表达,增强其对化疗药物顺铂的敏感性。     

Smac和XIAP与卵巢癌细胞耐药及凋亡关系的研究

目的探讨Smac与卵巢癌耐药及凋亡机制。方法采用RT-PCR、Western blot方法分别从基因和蛋白水平检测卵巢癌细胞系SKOV3及其耐药细胞系SKOV3/DDP中Smac与XIAP的表达水平的差异。构建Smac真核表达载体pc DNA3.1(+)/EGFP+Smac。采用脂质体转染法将pc DNA3.1(+)/EGFP+Smac真核表达载体导入卵巢癌顺铂耐药细胞系SKOV3/DDP。运用Western blot法检测转染前后细胞内Smac与XIAP表达的变化,MTT法检测Smac表达增加后细胞生长抑制率,观察Smac基因对卵巢癌细胞凋亡的影响。结果 SKOV3、SKOV3/DDP细胞系中均表达Smac/DIABLO与XIAP,且SKOV3中Smac的相对含量高于SKOV3/DDP。SKOV3中XIAP的相对含量低于SKOV3/DDP。Western blot检测转染后耐药细胞Smac蛋白表达增加,XIAP表达无明显改变。MTT结果转染48 h与72 h后细胞生长抑制率出现明显增加,差异有统计学意义(P<0.01)。结论 Smac表达的上调对卵巢癌细胞XIAP表达无明显影响,但对肿瘤细胞生长有明显的抑制作用。提示Smac确实促进卵巢癌细胞凋亡,但其凋亡机制并不是促进XIAP降解,而是与XIAP相互结合抑制其与caspase结合,激活caspase进而起到促凋亡作用。     

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.     

2-Deoxy-2-[F-18]fluoro-<Emphasis Type="SmallCaps">d</Emphasis>-glucose Accumulation in Ovarian Carcinoma Cell Lines

Purpose To evaluate 2-deoxy-2-[F-18]fluoro-d-glucose (FDG) accumulation in human ovarian carcinoma cell lines compared with control tumor cell lines known to accumulate FDG. Procedures FDG accumulation assays were performed in 15 different ovarian carcinoma cell lines at 1, 2, and 3 hours after incubation with 1 μCi of FDG. Results were compared with FDG accumulation in six different control tumor cell lines. 2-Deoxy-2-[F-18]fluoro-d-glucose accumulation was expressed as counts per minute (cpm) in cells and normalized to initial cpm in medium and total protein content of cell lysates. Results FDG accumulation in all 15 ovarian carcinoma cell lines was equal to or higher than 0.0005 ± 8.6 10⁻⁵ cpm in cells/cpm in medium/μg protein at all three different time points. In two ovarian carcinoma cell lines (ES-2, poorly differentiated clear cell carcinoma, and OVCAR-3, poorly differentiated papillary adenocarcinoma), FDG accumulation was not statistically, significantly different compared to the control cell line with the highest FDG accumulation (LS 174T human colorectal adenocarcinoma) at two or more time points (P ≥ 0.07). In 2 of 15 (13%) ovarian carcinoma cell lines (OVCAR5 epithelial carcinoma and SKOV3 clear cell carcinoma), FDG accumulation was lower than that in the control cell line with the lowest FDG accumulation (HT-29 human colorectal adenocarcinoma) at one or more time points (P < 0.05). Conclusions Most human ovarian carcinoma cell lines showed comparable FDG accumulations with control cell lines known to accumulate FDG. This study lays the foundations for further comparisons with other ovarian cancer cell lines and for other positron emission tomography tracers.     

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.     

Doxorubicin loaded ferritin nanoparticles for ferroptosis enhanced targeted killing of cancer cells

In recent years, ferroptosis has been investigated widely as a new form of cell death. Development of nanodrugs for ferroptosis induction in cancer cells may be a promising approach for cancer treatment. Here, we developed a type of nanoparticle consisting of the antitumor drug doxorubicin and exogenous ferritin. The drug loading process did not change the size of ferritin obviously. And this nanoparticle could induce the accumulation of ROS and cell ferroptosis for transferrin receptor overexpressed tumor cell, HT29. The ferroptosis process was also confirmed using inhibitors for ferroptosis. The cytotoxicity of this nanoparticle is similar to that of free DOX. This study provides a new strategy for targeting and killing transferrin receptor overexpressed tumor cells.

DOX loaded ferritin selectively induces ferroptosis enhanced killing of transferrin receptor 1 overexpressed cancer cells.     

Ber-EP4 and anti-calretinin antibodies: a useful combination for differential diagnosis of various histological types of ovarian cancer cells and mesothelial cells

The differential diagnosis between reactive mesothelial cells and ovarian carcinoma cells is often difficult in cytologic specimens. Immunocytochemical procedures have been utilized in assisting this differential diagnosis, with limitations. Furthermore, previous studies examined only serous type but not other histological types of ovarian carcinoma cases. Therefore, we evaluated the practical value of various epithelial and mesothelial markers in differential diagnosis of these two types of cells. Various types of ovarian carcinoma (serous, n = 22; mucinous, n = 10; endometrioid, n = 7; clear cell, n = 10) and benign mesothelial tissues (n = 15) were studied by immunohistochemistry. We then studied effective panels of antibodies by immunohistochemistry in 43 cytologic specimens of ascites or peritoneal lavage fluid consisting of 20 reactive mesothelium and 23 adenocarcinomas of the ovary. In the tissue specimens, Ber-EP4, a monoclonal antibody of epithelial antigen, and a polyclonal antibody against calretinin, which is expressed in mesothelium, are used in differentiating reactive mesothelial cells from ovarian carcinoma. In cytologic specimens, the sensitivity and specificity of Ber-EP4 were 100% and 90%, respectively. The sensitivity and specificity of the anti-calretinin antibody were 90% and 91%, respectively. Using multiple regression analysis, the correlation coefficient between epithelial antigen and calretinin reactivity was r = 0.938, with a significance level of p < 0.0001. In conclusion, the combined immunostaining of cytologic specimens for Ber-EP4 and the anti-calretinin antibody is helpful for the differential diagnosis between mesothelial cells and not only serous type, but also mucinous, endometrioid and clear cell types of ovarian cancer cells.     

Soluble CD40 ligands sensitize the epithelial ovarian cancer cells to cisplatin treatment

ObjectiveCD154 (CD40L) is a protein that is primarily expressed on activated T cells and is a member of the TNF superfamily of molecules. It binds to CD40 on antigen-presenting cells (APC), which leads to many effects depending on the target cell type. Being an activator of immune cells, CD40L has also been shown to directly induce apoptosis in tumor cells by multiple mechanisms. To understand the role of sCD40L in regulating the proliferation of epithelial ovarian cancer cells treated or untreated with cisplatin.MethodsEpithelial ovarian cancer cells: SKOV3 and its cisplatin-resisitant strain SKOV3/DDP cells were used to test the effect of sCD40L and cisplatin. The proliferation of SKOV3 and SKOV3/DDP cells were measured by MTT. Cell cycle was assessed by flow cytometry. The mRNA expressions of targeted genes were detected by qRT-PCR. The protein expressions were detected by Western blotting.ResultssCD40L showed a significant dose-dependence inhibitory effect on the proliferation of ovarian cancer cell lines. sCD40L in combination with cisplatin could sensitized SKOV3/DDP cells to cisplatin treatment and reversed the drug resistance of SKOV3/DDP cells. The reversal ratios of 1 μg/ml sCD40L combined with cisplatin in SKOV3 and SKOV3/DDP cells were 2.11, 2.71, while the reversal ratios of 2 μg/ml sCD40L combined with cisplatin in SKOV3 and SKOV3/DDP cells were 3.78, 5.20, respectively. sCD40L or sCD40L combined cisplatin increased tumor cells in G0/G1 phase. sCD40L in combination with cisplatin decreased the expression levels of GST-π, LRP, Survivin, p53 and Bcl-2 in both epithelial ovarian cancer cell lines. The protein expression level of GST-π, LRP and P53 protein was also decreased upon sCD40L in combination with cisplatin although the expression level of Bcl-2 and survivin protein had no significant difference.ConclusionsCD40L inhibits the proliferation of SKOV3 and SKOV3/DDP cells. The combined application of sCD40L and cisplatin can strength the inhibitory effect of cisplatin, and to a certain extent, reversing the resistance to cisplatin in SKOV3/DDP cells. sCD40L could lead a cell block in G0/G1 phase and make the cell growth restrained. sCD40L could induce SKOV3 and SKOV3/DDP cells apoptosis and reverse drug resistance through cutting GST-π mRNA, LRP mRNA, survivin mRNA, p53 mRNA and Bcl-2 mRNA and decreasing the expression of GST-π, LRP and P53 protein in SKOV3 and SKOV3/DDP cells, which provides in-vivo experiment basis to the application of sCD40L as a drug improving ovarian cancer cells sensitivity to cisplatin.     

Erythropoietin inhibits apoptosis induced by photodynamic therapy in ovarian cancer cells

Recombinant human erythropoietin is widely used to treat anemia associated with cancer and with the myelosuppressive effects of chemotherapy, particularly platinum-based regimens. Erythropoietin is the principal regulator of erythroid cell proliferation, differentiation, and apoptosis. Recently, the antiapoptotic and proliferative effects of erythropoietin on nonhematopoietic cells were also established. We now show the effect of erythropoietin treatment on the response of A2780 and SKOV3 ovarian carcinoma cell lines to photodynamic therapy (PDT) using hypericin. SKOV3 exhibited an increased resistance to hypericin when cells were treated with erythropoietin. This resistance was reversed by treatment of SKOV3 cells with the specific Janus kinase 2 kinase inhibitor AG490 or the tyrosine kinase inhibitor genistein. These results support a role for the specific erythropoietin-induced Janus kinase 2/STAT signal transduction pathway in PDT resistance. Evidence of erythropoietin signaling was obtained by the demonstration of Akt phosphorylation in both A2780 and SKOV3 cells. Erythropoietin-treated SKOV3 cells exhibited decreased apoptosis induced by hypericin, an effect that was blocked by the phosphoinositide 3-kinase/Akt inhibitor wortmannin. These results may have important implications for ovarian cancer patients undergoing PDT and receiving erythropoietin.     

Virus directed enzyme prodrug therapy for ovarian and pancreatic cancer using retrovirally delivered E. coli nitroreductase and CB1954.

Expression of the E. coli enzyme nitroreductase (NTR) in tumour cells enables them to activate the prodrug CB1954 (5-(aziridin-1-yl)-2,4-dinitrobenzamide), leading to interstrand DNA crosslinking and cell death. Using transfected or retrovirally transduced SKOV3 ovarian carcinoma cell clones, we show a strong correlation between sensitivity to CB1954 and level of NTR enzyme activity. Importantly for clinical application in ovarian cancer, a cisplatin-resistant ovarian tumour cell line remains as susceptible to the NTR-dependent cytotoxicity of CB1954 as parental cells. In mixed populations of NTR-expressing and non-expressing cells, we observe a marked 'bystander killing' effect with this system. The use of NTR-encoding retroviruses from clonal producer cell lines at titres of 5 x 10(5) c.f.u./ml to transduce either established or low passage primary ovarian carcinoma lines only achieves an average 10-fold sensitisation of the cultures at gene transfer efficiencies of 15-25%. Concentration of the retrovirus to 3 x 10(7) c.f.u./ml elevates gene transfer to 80-90% in a single exposure to target cells, resulting in up to 500-fold sensitisation of the entire, unselected SKOV3 population to CB1954. In an initial investigation of NTR/CB1954 for the treatment of tumours in vivo, we observe regression of tumours expressing NTR following administration of CB1954, resulting in significantly increased median survival.     

MiR-139-5p通过靶向NFAT抑制卵巢癌SKOV3细胞的生长、集落形成、侵袭和迁移

目的:探究miR-139-5p对卵巢癌SKOV3细胞的生长、集落形成、侵袭能力以及迁移能力的影响及其机制。方法:采用qRT-PCR检测卵巢癌患者癌组织和卵巢癌细胞SKOV3中miR-139-5p表达情况。MiR-139-5p mimic转染SKOV3细胞,WST比色实验、集落形成实验和Transwell实验分别检测细胞的活性、集落形成、侵袭和迁移能力。采用TargetScan在线软件筛选miR-139-5p的潜在靶基因NFAT,并进一步验证。结果:MiR-139-5p在卵巢癌组织和SKOV3细胞中表达异常降低。MiR-139-5p过表达显著抑制SKOV3细胞的生长、集落形成、迁移及侵袭能力。NFAT是miR-139-5p的靶基因。过表达NFAT能逆转miR-139-5p过表达对SKOV3细胞集落形成、侵袭能力以及迁移能力与侵袭的抑制作用。结论:MiR-139-5p通过抑制靶基因NFAT来抑制卵巢癌SKOV3细胞的生长、集落形成、侵袭和迁移能力。     

负载卵巢癌冻融抗原对树突状细胞分泌细胞因子的影响

目的探讨负载卵巢癌冻融抗原对树突状细胞分泌细胞因子的影响。方法采用反复冻融法获得卵巢癌细胞株SKOV3细胞抗原,联合应用重组人粒细胞-巨噬细胞集落刺激因子、重组人白细胞介素-4和重组人肿瘤坏死因子-α,体外诱导人外周血CD14^＋单核细胞为成熟DCs并负载卵巢癌肿瘤抗原;采用ELISA法检测培养细胞上清液中分泌IL-12p70、IFN-γ和IL-10含量,评价非成熟DCs和成熟DCs以及肿瘤抗原负载DCs和未负载肿瘤抗原DCs激活的CTL分泌细胞因子的能力。结果联合应用重组人的GM-CSF、IL-4和TNF-α可在体外诱导出成熟DCs;经ELISA方法检测,不同状态下的DCs分泌IL-12p70、IFN-γ的量不同,成熟DCs较非成熟DCs有更强的分泌IL-12p70、IFN-γ的能力（P〈0.05）,负载抗原DCs激活的CTL较未负载抗原DCs激活的CTL有更强的分泌IL-12p70[（182.89±4.57）pg/ml和（99.76±5.42）pg/ml,P〈0.01]和IFN-γ的能力[（102.11±5.95）pg/ml和（68.29±5.04）pg/ml,P〈0.01]。而成熟DCs分泌IL-10的能力与非成熟DCs相比,差异无统计学意义（P〉0.05）。负载抗原后DCs激活的CTL与未负载抗原DCs激活CTL上清液中IL-10的浓度相比,两者之间的差异无统计学意义（P〉0.05）。结论细胞因子的分泌量受DCs的成熟状态及某些刺激信号的影响,卵巢癌冻融抗原是其刺激信号之一。     

Long non-coding RNA NEAT1 regulates ferroptosis sensitivity in non-small-cell lung cancer

ObjectivesFerroptosis is caused by iron-dependent lipid peroxide accumulation, the sensitivity of which might be regulated by acyl-CoA synthetase long chain family member 4 (ACSL4). Non-small-cell lung cancer (NSCLC) can resist oxidative stress and reduce the sensitivity of tumor cells to ferroptosis by changing the expression of some proteins. Mechanisms involving ferroptosis sensitivity in NSCLC are not fully understood.MethodsA dual-luciferase reporter assay was used to confirm a targeting relationship between long non-coding (lnc)RNA NEAT1 and ACSL4. Overexpression and silencing assays of NEAT1 function were used to determine its roles in cell death (by TUNEL staining) and lipid peroxidation (by malondialdehyde levels). Expression of ferroptosis-related proteins (SLCA11, GPX4, and TFR4) was evaluated by western blot in NSCLC cells treated or not with the ferroptosis inducer erastin.ResultsErastin-induced cell death was positively correlated with ACSL4 level. NEAT1 regulated levels of ACSL4 and proteins related to the ferroptosis and classical apoptosis pathways. Levels of ACSL4, SLC7A11, and GPX4 were decreased more by NEAT1 silencing plus erastin than by erastin alone.ConclusionNEAT1 regulates ferroptosis and ferroptosis sensitivity, with the latter depending on ACSL4, suggesting that targeting NEAT1 or ACSL4 may be a viable therapeutic approach to the treatment of NSCLC.     

超声引导下细针穿刺细胞学检查在妇科盆腔肿瘤诊治中的应用

目的:探讨超声引导下细针穿刺细胞学(fine needle aspiration,FNA)检查对妇科盆腔肿瘤的临床诊断与治疗价值。方法:32例妇科盆腔肿瘤患者在B超引导下进行FNA检查,同时送组织病理学检查。结果:32例盆腔肿瘤患者中,FNA检查诊断为卵巢恶性肿瘤28例,盆腔良性病变4例。FNA诊断为恶性肿瘤的28例患者中,除3例因针吸样本量较少未能作出组织病理学诊断外,余25例组织病理学诊断为浆液性、黏液性、乳头状癌或子宫内膜样腺癌等。FNA诊断为良性肿瘤的4例患者中,术后组织病理学证实为卵巢畸胎瘤、卵巢浆液性囊腺瘤、子宫肌瘤、附件炎性反应。FNA检查对盆腔肿物诊断的敏感性和特异性均为100.0%。32例患者均未发现针道种植及感染。结论:FNA诊断妇科肿瘤的敏感性及特异性高,具有重要的临床价值。     

Inhibition activity of sulfated polysaccharide of Sepiella maindroni ink on matrix metalloproteinase (MMP)-2.

SIP-SII is the sulfated S. maindroni ink polysaccharide (SIP) isolated from cuttlefish Sepiella maindroni. SIP-SII weakly inhibited tumor cell growth without cytotoxicity in vitro assay. Herein, we examined the effects of SIP-SII on the expression of matrix metalloproteinase MMP-2 and MMP-9 as well as tumor cell invasion and migration. SIP-SII (0.8-500 microg/ml) significantly decreased the expression of MMP-2 activity in human ovarian carcinoma cells SKOV3 as evidenced by the gelatin zymography analysis. No significant decrease of MMP-9 was detected in the cell line after SIP-SII treatment. The expression of MMP-2 was also evaluated using Western blot analysis. The results showed that SIP-SII inhibited the expression of MMP-2 in SKOV3 and human umbilical vein vascular endothelial cells ECV304 after 24 h incubation. Furthermore, the activity of invasion and migration of SKOV3 and ECV304 cells were measured. SIP-SII displayed an inhibitory effect on the penetration of SKOV3 cells through Matrigel-coated membrane in transwell chamber. A significant inhibition of ECV304 cell migration was observed in the presence of SIP-SII. These results suggest that SIP-SII might suppress invasion and migration of carcinoma cells via inhibition of MMP-2 proteolytic activity.