Shikonin from Chinese herbal medicine induces GSDME-controlled pyroptosis in tumor cells

ObjectiveTo investigate the potential anti-tumor mechanisms of naphthoquinone compound shikonin (SKN) extracted from the root of Chinese herbal medicine plant lithospermum (Lithospermum erythrorhizon Sieb. & Zucc.).MethodsWe first observed that SKN treatment led to swelling and bubbles in HeLa cells that were similar to the phenotype of cell pyroptosis. Subsequently, the HeLa cells experienced a pyroptotic process with SKN, and this was then assessed using lactate dehydrogenase (LDH) release and propidium iodide (PI)/Hoechst double staining experiments. Pyroptosis is defined as gasdermin-mediated programmed necroptosis. To identify the potential pyroptosis machinery, two strategies were utilized that included a genome-wide clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 screening experiment and a pyroptosis reconstitution assay executed by each of the five known gasdermins (GSDMA-E). Moreover, endogenous cleavage was also detected in a panel of tumor cell lines.ResultsCompared with the control, both the LDH release and PI/Hoechst double-staining experiments suggested that SKN induced perforation and enhancement of the permeability of the cell membranes that resulted in pyroptosis in HeLa cells (P = .028 and P = .032, respectively). In addition, the reconstitution assays in human embryonic kidney 293T (HEK-293T) cells and endogenous cleavage assays in HeLa cells indicated that the pyroptosis was controlled by GSDME. In addition, we also found SKN could trigger pyroptosis in a panel of tumor cell lines in which the cellular morphologies were proportional to the GSDME expression levels. Additionally, the cleavage of GSDME was also detected, and this was indicative of a similar GSDME-mediated mechanism.ConclusionOur study not only explained the molecular mechanism of cytotoxicity of SKN to various tumor cells, but also provided additional information for the potential clinical application of natural naphthoquinone compounds against cancer.     

Immunotherapy in Hodgkin and non-Hodgkin lymphoma: Innate,adaptive and targeted immunological strategies

Since the clinical introduction of anti-CD20 monoclonal antibodies into lymphoma treatment, immunologic approaches in lymphoma have made substantial progress. Advances in our understanding of tumor immunology have led to the development of strategies to overcome immunologic barriers responsible for an ineffective immune response. Specifically, therapeutic agents have been developed and tested against molecules that are responsible for T-cell exhaustion. The use of monoclonal antibodies against immune checkpoints in the adaptive immune system, such as programmed cell death-1 and cytotoxic T-lymphocyte-associated protein 4, has changed the landscape of cancer therapy including the treatment of lymphoma. This achievement has recently been accompanied by the development of novel immune checkpoint inhibitors targeting the innate immune system, including the CD47-SIRPα signaling pathway, and this approach has yielded promising results. To overcome impaired antigen presentation, antibody-based cytotoxic strategies, namely antibody-drug conjugates (polatuzumab vedotin and brentuximab vedotin) and bispecific T-cell or NK-cell engagers (blinatumomab, REGN1979, RG6206, and AFM13), have rapidly evolved with promising clinical activity. As additional tools become available for lymphoma treatment, formulation of safe, rational combination strategies to combine them with standard therapy will be of paramount importance. A successful approach to the treatment of lymphoma may require both an optimized anti-tumor immune response as well as effective depletion of malignant lymphoid cells.     

黄连素对人胃癌细胞SGC7901凋亡影响的研究

目的:探讨黄连素对人胃癌细胞SGC7901凋亡的影响。方法:MTS法检测不同浓度的黄连素(100、150、200μmol/L)对胃癌细胞的抑制作用,Hoechst 33258染色检测不同浓度的黄连素(100、150、200μmol/L)对细胞凋亡的影响;Real Time Q-PCR检测胃癌细胞中Cleaved Caspase-3、Bcl-2、Bax的mRNA表达;Western blot检测胃癌细胞中Cleaved Caspase-3、Bcl-2、Bax的蛋白表达。结果:不同浓度的黄连素能显著降低人胃癌细胞SGC701活性(P<0.05,P<0.01),促进其凋亡,升高Cleaved Caspase-3、Bax的mRNA和蛋白表达水平(P<0.05,P<0.01),降低Bcl-2的mRNA和蛋白表达水平(P<0.05,P<0.01)。结论:不同浓度的黄连素可诱导胃癌细胞凋亡,其机制可能与升高Cleaved Caspase-3、Bax的表达,降低Bcl-2的表达有关。     

Management of hepatitis B reactivation in immunosuppressed patients: An update on current recommendations

The proportion of hepatitis B virus(HBV) previously exposed patients who receive immunosuppressive treatment is usually very small. However, if these individuals are exposed to potent immunosuppressive compounds, the risk of HBV reactivation(HBVr) increases with the presence of hepatitis B surface antigen(HBsAg) in the serum. Chronic HBsAg carriers have a higher risk than those who have a total IgG anticore as the only marker of resolved/occult HBV disease. The loss of immune control in these patients may results in the reactivation of HBV replication within hepatocytes. Upon reconstitution of the immune system, infected hepatocytes are once again targeted and damaged by immune surveillance in an effort to clear the virus. There are different virological scenarios, and a wide spectrum of associated drugs with specific and stratified risk for the development of HBVr. Some of this agents can trigger a severe degree of hepatocellular damage, including hepatitis, acute liver failure, and even death despite employment of effective antiviral therapies. Currently, HBVr incidence seems to be increasing around the world; a fact mainly related to the incessant appearance of more powerful immunosuppressive drugs launched to the market. Moreover, there is no consensus on the length of prophylactic treatment before the patients are treated with immunosuppressive therapy, and for how long this therapy should be extended once treatment is completed. Therefore, this review article will focus on when to treat, when to monitor, what patients should receive HBV therapy, and what drugs should be selected for each scenario. Lastly, we will update the definition, risk factors, screening, and treatment recommendations based on both current and different HBV management guidelines.     

The neglected brothers come of age: B cells and cancer

The opposing roles of innate and adaptive immune cells in suppressing or supporting cancer initiation, progression, metastasis and response to therapy has been long debated. The mechanisms by which different monocyte and T cell subtypes affect and modulate cancer have been extensively studied. However, the role of B cells and their subtypes have remained elusive, perhaps partially due to their heterogeneity and range of actions. B cells can produce a variety of cytokines and present tumor-derived antigens to T cells in combination with co-stimulatory or inhibitory ligands based on their phenotype. Unlike most T cells, B cells can be activated by innate immune stimuli, such as endotoxin. Furthermore, the isotype and specificity of the antibodies produced by plasma cells regulate distinct immune responses, including opsonization, antibody-mediated cellular cytotoxicity (ADCC) and complement activation. B cells are shaped by the tumor environment (TME), with the capability to regulate the TME in return. In this review, we will describe the mechanisms of B cell action, including cytokine production, antigen presentation, ADCC, opsonization, complement activation and how they affect tumor development and response to immunotherapy. We will also discuss how B cell fate within the TME is affected by tumor stroma, microbiome and metabolism.     

Solid extraintestinal malignancies in patients with inflammatory bowel disease

Malignancies constitute the second cause of death in patients with inflammatory bowel diseases (IBD), after cardiovascular diseases. Although it has been postulated that IBD patients are at greater risk of colorectal cancer compared to the general population, lately there has been evidence supporting that this risk is diminishing over time as a result of better surveillance, while the incidence of extraintestinal cancers (EICs) is increasing. This could be attributed either to systemic inflammation caused by IBD or to long-lasting immunosuppression due to IBD treatments. It seems that the overall risk of EICs is higher for Crohn’s disease patients and it is mainly driven by skin cancers, and liver-biliary cancers in patients with IBD and primary sclerosing cholangitis. The aims of this review were first to evaluate the prevalence, characteristics, and risk factors of EICs in patients with IBD and second to raise awareness regarding a proper surveillance program resulting in early diagnosis, better prognosis and survival, especially in the era of new IBD treatments that are on the way.     

乌头碱调节 C-C基序趋化因子配体 2/C-C基序趋化因子受体 2信号通路对膀胱癌细胞抗肿瘤活性及其作用机制研究

目的探讨乌头碱调节 C-C基序趋化因子配体 2(CCL2)/C-C基序趋化因子受体 2(CCR2)信号通路对膀胱癌细胞抗肿瘤活性及其作用机制研究。方法研究起止时间为 2021年 12月至 2022年 10月。采用细胞计数试剂盒( CCK-8)法检测 2.5、5.0、10.0、20.0、30.0 μmol/L乌头碱处理后的人膀胱癌 5637细胞存活率,筛选出合适的乌头碱作用浓度。将 5637细胞分为对照组、乌头碱低剂量( 10 μmol/L)组、乌头碱高剂量( 20 μmol/L)组、乌头碱高剂量(20 μmol/L)+空载组、乌头碱高剂量( 20 μmol/L) +CCL2过表达组,分组处理后,采用 CCK-8法、 5-乙炔基 -2''-脱氧尿苷( Edu)染色法及 Hoechst 33258染色分别检测各组 5637细胞存活率、增殖率、凋亡率;采用 Transwell实验及划痕实验分别检测各组 5637细胞侵袭数、迁移率;采用免疫荧光染色检测各组 5637细胞凋亡相关蛋白 BCL2相关 X蛋白( Bax)和 B淋巴细胞瘤 -2(Bcl-2)表达比值( Bax/Bcl-2);采用免疫印迹法检测各组 5637细胞 CCL2/CCR2信号通路相关蛋白及上皮间质转化标志蛋白［神经钙黏素( N-cadherin)、锌指 E盒结合同源盒 1(ZEB1)、紧密连接蛋白 1(ZO-1)、上皮钙黏素( E-cadherin)］表达。结果与对照组相比,乌头碱低剂量组、乌头碱高剂量组、乌头碱高剂量 +空载组细胞 CCL2(0.69±0.09、0.20±0.03、0.19±0.04比 1.21±0.13)CCR2(0.78±0.12、0.26±0.06、0.27±0.07比 1.33±0.20)、 Ncadherin(0.65±0.06、0.12±0.02、0.11±0.03比 1.24±0.12)与 ZEB1蛋白存活率、增殖率、侵袭数、迁移率均降低( P<0.05), Bax/Bcl-2、细胞 ZO-1与 E-cadherin蛋白表达均升高( P<0.05);乌头碱高剂量组、乌头碱高剂量 +空载组细胞 CCL2、CCR2、N-cad? herin与 ZEB1蛋白表达、存活率、增殖率、侵袭数、迁移率相比乌头碱低剂量组进一步降低( P<0.05)Bax/Bcl-2、细胞 ZO-1与 Ecadherin蛋白表达进一步升高( P<0.05)。与乌头碱高剂量组相比,乌头碱高剂量 +CCL2过表达组CCL2、CCR2、N-cadherin与 ZEB1蛋白表达、存活率、增殖率、侵袭数、迁移率升高( P<0.05)Bax/Bcl-2、细胞 ZO-1与 E-cadherin蛋白表达降低( P<0.05)。结论乌头碱可通过 CCL2/CCR2信号通路而抑制膀胱癌细胞存增殖及侵袭和迁移,促使其凋亡。     

鹅去氧胆酸浙贝乙素酯对H22荷瘤小鼠抑瘤作用*

目的研究新型化合物鹅去氧胆酸浙贝乙素酯(CDCA-Ver)对H22荷瘤小鼠体内肿瘤细胞生长和免疫器官的影响。方法采用鼠系肝癌细胞H22造荷瘤模型,将H22荷瘤小鼠40只,采用随机数字表法随机分成4组,每组10只,分别为模型对照组、环磷酰胺(CTX)组、CDCA-Ver腹腔注射组和CDCA-Ver静脉注射组。模型对照组按每日10 mL·kg-1无菌0.9%氯化钠溶液腹腔注射1次,CTX组按每日20 mg·kg-1剂量腹腔注射1次,CDCA-Ver腹腔注射组每日按20 mg·kg-1剂量腹腔注射1次,CDCA-Ver静脉注射组每日按20 mg·kg-1剂量小鼠尾静脉注射1次。接种24 h后给药,给药量为0.1 mL·(10 g)-1,连续给药10 d。以CTX为阳性对照药,考察CDCA-Ver(静脉注射和腹腔注射)对荷瘤小鼠肿瘤的生长抑制作用,评价CDCA-Ver对荷瘤小鼠的免疫脏器指数(胸腺指数和脾指数)的影响,采用组织病理学切片法研究CDCA-Ver对荷瘤小鼠瘤块组织病理学形态的影响。结果CDCA-Ver静脉注射和腹腔注射两种给药方式对荷瘤小鼠的肿瘤生长均具有良好的抑制作用,腹腔注射20 mg·kg-1剂量CDCA-Ver抑瘤率达48.3%,与模型对照组比较差异有统计学意义(P＜0.05),与CTX组作用相当。与模型对照组相比,CDCA-Ver腹腔注射组脾指数和胸腺指数没有显著变化(P＞0.05),而CTX组脾指数和胸腺指数均显著降低(P＜0.01),表明CDCA-Ver发挥体内抗肿瘤作用不会降低荷瘤小鼠的免疫功能。组织病理学结果也证实CDCA-Ver具有体内抗肿瘤作用。结论CDCA-Ver对H22荷瘤小鼠肿瘤生长具有明显的抑制作用。     

抗肿瘤多肽类药物的来源及其机制的研究进展

恶性肿瘤已成为严重影响人类健康的一种疾病,其治疗手段一直是人们所探索的内容,药物治疗就是其中一种治疗方法。与传统化疗药物相比,抗肿瘤多肽类药物以其分子量小、特异性强、毒性低等特点作为新的治疗肿瘤药物一直广受人们关注。多肽的来源广泛,有存在于天然动植物及微生物体内,也可以通过蛋白质酶解或人工合成得到。近年来,越来越多的多肽类药物被发现除抗菌外还具有抗肿瘤的作用,其抗肿瘤的机制多种多样但尚未完全清楚,是许多研究者研究的重点。本文从多肽类药物的来源与特点及它们的抗肿瘤机制等方面对多肽类药物的研究进展进行综述。