PD-1/PD-L1抑制剂联合传统治疗手段治疗晚期非小细胞肺癌的研究进展

肺癌是全球发病率及死亡率极高的一种恶性肿瘤。非小细胞肺癌(NSCLC)约占所有肺癌的85%以上。以往针对NSCLC的治疗手段主要包括手术、化疗、放疗和靶向治疗。近年来,对于肺癌在内的多种实体瘤的治疗,免疫检查点抑制剂的免疫治疗取得了显著进展,越来越多的临床试验肯定了免疫治疗联合传统治疗手段在NSCLC中的疗效。本文将着重介绍程序性死亡蛋白1/程序性死亡蛋白配体1抑制剂联合传统治疗手段在治疗NSCLC中的研究进展。     

免疫检查点及PD-1/PD-L1抑制药在肿瘤免疫诊断治疗中的应用

程序性死亡受体-1(PD-1)/程序性死亡配体-1(PD-L1)单克隆抗体为代表的免疫检查点抑制药在肿瘤治疗领域取得了巨大的成功。故对免疫检查点抑制药在肿瘤免疫中的作用进行概述,并分析了免疫检查点抑制药的研究热点PD-1/PD-L1通路在抗肿瘤免疫的治疗意义,再对目前PD-1/PD-L1抑制药在诊断、治疗和预后中的应用调查,在此基础上分析PD-1/PD-L1抑制药进一步应用需要研究的问题,以期为PD-1/PD-L1抑制药进一步应用提供研究参考。     

程序性死亡配体-1在人非小细胞肺癌组织中的表达及临床意义

目的：探讨程序性死亡配体‐1（PD‐L1）在人非小细胞肺癌（NSCLC）组织中的表达及临床意义。方法采用Western blot和免疫组织化学染色法检测41例NSCLC患者的肺癌组织、癌旁组织、正常肺组织和15例肺部良性病变患者的正常肺组织中的 PD‐L1表达,分析NSCLC患者PD‐L1表达的影响因素。结果 NSCLC患者肺癌组织中 PD‐L1阳性表达率为80.49％（33／41）,肺良性病变患者正常肺组织中几乎无PD‐L1表达。NSCLC患者肺癌组织中 PD‐L1表达高于癌旁组织及正常肺组织（P＜0．05）。PD‐L1的表达与肿瘤患者分期、淋巴结转移和远处转移相关（P＜0．05）,而与肿瘤病理分型、患者年龄、性别、吸烟史及肿瘤大小等无关（ P＞0．05）。结论 PD‐L1在NSCLC组织中显著高表达,在NSCLC进展中发挥重要作用,可能成为NSCLC预后的判断指标及免疫治疗的新靶点。     

二线化疗联合PD-1/PD-L1抑制剂治疗晚期非小细胞肺癌的临床疗效分析

目的 分析二线化疗联合程序性死亡受体1/程序性死亡-配体1(PD-1/PD-L1)抑制剂治疗晚期非小细胞肺癌的临床疗效。方法 48例晚期非小细胞肺癌患者,根据随机数字表法分为对照组和研究组,每组24例。两组均接受注射用紫杉醇(白蛋白结合型)+顺铂静脉注射治疗,对照组联合安慰剂治疗,研究组联合PD-1/PD-L1抑制剂治疗。比较两组治疗效果以及毒副反应发生情况。结果 研究组治疗总有效率62.50%高于对照组的33.33%,差异具有统计学意义(P<0.05)。研究组胃肠道反应、肝肾异常、血小板减少、白细胞减少、骨髓抑制发生率分别为25.00%、12.50%、8.33%、20.83%、12.50%,与对照组的33.33%、16.67%、12.50%、20.83%、20.83%比较,差异无统计学意义(P>0.05)。结论 对晚期非小细胞癌患者采用二线化疗联合PD-1/PD-L1抑制剂治疗,可提高治疗效果,且毒副反应相对较低,临床价值明确。     

肿瘤免疫检查点PD-1/PD-L1抑制剂的中药活性成分研究进展

肿瘤免疫治疗已成为新型的癌症治疗手段,有望彻底消除肿瘤。免疫检查点抑制剂,特别是程序性死亡受体-1（PD-1）和程序性死亡受体-配体1（PD-L1）抗体在多种实体瘤的治疗中取得很好的临床疗效,但是生物制剂存在免疫原性强、价格昂贵等缺点,因此,寻找免疫检查点小分子抑制剂成为未来肿瘤免疫疗法的新挑战。本文将综述近年发现的抑制PD-1/PD-L1表达的中药活性小分子及其对肿瘤免疫微环境的调控作用。     

五味子丙素抑制脂多糖诱导心肌细胞的炎症反应与 细胞焦亡的作用及机制研究

目的：探究五味子丙素（SchC）对脂多糖（LPS）诱导心肌细胞HL-1炎症反应与细胞焦亡的影响。方法：培养小鼠心肌细胞HL-1,将其分为空白对照组、模型组（LPS）、LPS＋SchC组。SchC预处理1 h后,分别以LPS刺激24 h或48 h,ELISA法提取细胞培养液上清液,检测细胞因子IL-1β、HMGB1和TNF-α的分泌,MTT法收集细胞检测细胞活力,Western Blot检测cleaved-Caspase1、GSDMD-N和NLRP3的表达;将HL-1细胞分为空白对照组、模型组（LPS）、LPS＋siGSDMD（GSDMD siRNA）组、LPS＋siGSDMD+SchC组,给药预处理1 h后,LPS刺激24 h,收集细胞培养液上清液,ELISA法检测IL-1β的分泌,MTT法检测细胞活力。结果：与LPS组比较,LPS＋SchC组细胞活力得到改善（P<0.05）,IL-1β、HMGB1和TNF-α的分泌均显著降低（P<0.05）;Western Blot结果表明,与LPS组比较,LPS＋SchC组的cleaved-Caspase1、GSDMD-N段和NLRP3蛋白水平显著降低（P<0.05）;ELISA和MTT结果表明,与LPS组比较,LPS＋siGSDMD组IL-1β的分泌显著降低（P<0.05）且细胞活力显著得到改善（P<0.01）,然而LPS＋siGSDMD组与LPS＋siGSDMD+SchC组的IL-1β分泌水平和细胞活力并无显著性差异。结论：五味子丙素能有效缓解LPS诱导HL-1的炎症反应和细胞焦亡。     

五味子丙素抑制脂多糖诱导心肌细胞的炎症反应与 细胞焦亡的作用及机制研究

目的:探究五味子丙素(SchC)对脂多糖(LPS)诱导心肌细胞HL-1炎症反应与细胞焦亡的影响。方法:培养小鼠心肌细胞HL-1,将其分为空白对照组、模型组(LPS)、LPS+SchC组。SchC预处理1 h后,分别以LPS刺激24 h或48 h,ELISA法提取细胞培养液上清液,检测细胞因子IL-1β、HMGB1和TNF-α的分泌,MTT法收集细胞检测细胞活力,Western Blot检测cleaved-Caspase1、GSDMD-N和NLRP3的表达;将HL-1细胞分为空白对照组、模型组(LPS)、LPS+siGSDMD(GSDMD siRNA)组、LPS+siGSDMD+SchC组,给药预处理1 h后,LPS刺激24 h,收集细胞培养液上清液,ELISA法检测IL-1β的分泌,MTT法检测细胞活力。结果:与LPS组比较,LPS+SchC组细胞活力得到改善(P<0.05),IL-1β、HMGB1和TNF-α的分泌均显著降低(P<0.05);Western Blot结果表明,与LPS组比较,LPS+SchC组的cleaved-Caspase1、GSDMD-N段和NLRP3蛋白水平显著降低(P<0.05);ELISA和MTT结果表明,与LPS组比较,LPS+siGSDMD组IL-1β的分泌显著降低(P<0.05)且细胞活力显著得到改善(P<0.01),然而LPS+siGSDMD组与LPS+siGSDMD+SchC组的IL-1β分泌水平和细胞活力并无显著性差异。结论:五味子丙素能有效缓解LPS诱导HL-1的炎症反应和细胞焦亡。     

米诺环素治疗慢性牙周炎的临床疗效及其对外周血T细胞亚群、PD-1、PD-L1表达的影响

目的 探究米诺环素治疗慢性牙周炎的临床疗效及其对外周血T细胞亚群及程序性死亡分子-1（PD-1）、程序性死亡分子-1配体（PD-L1）表达的影响。方法 选择2015年1月-2017年1月在榆林市星元医院口腔科接受治疗的96例慢性牙周炎患者为研究对象。根据随机数字表法将患者分为对照组和观察组,每组各48例。对照组患者采用龈下刮治及根面平整进行治疗。观察组患者在对照组治疗的基础上在牙周袋底部缓慢注入盐酸米诺环素软膏,0.2 g/次,1次/周,边注射边后退。两组均治疗4周。观察两组患者的临床疗效,同时比较两组治疗前后的牙龈指数（GI）、菌斑指数（PLI）、牙槽骨吸收、附着丧失（CAL）以及牙周袋深度（PD）、C反应蛋白（CRP）、白细胞介素-10（IL-10）、龈沟出血指数（SBI）、外周血T淋巴细胞亚群和表面PD-1、PD-L1的表达情况。结果 治疗后,对照组患者临床总有效率为72.92%,显著低于观察组的91.67%（P<0.05）。治疗后,两组患者GI、PLI、PD、CAL和牙槽骨吸收等指标均显著降低（P<0.05）;且观察组以上各指标均显著低于对照组（P<0.05）。治疗后,两组患者龈沟液中CRP水平和SBI评分均显著降低,而龈沟液中IL-10水平显著升高（P<0.05）;且观察组龈沟液中CRP、IL-10水平和龈沟出血指数均优于对照组（P<0.05）。治疗后,两组患者外周血T淋巴细胞亚群中CD4⁺和CD4⁺/CD8⁺均显著降低,CD8⁺显著升高（P<0.05）;且观察组外周血T淋巴细胞亚群中CD4⁺和CD4⁺/CD8⁺均显著低于对照组,CD8⁺比例高于对照组（P<0.05）。治疗后,两组患者外周血CD4⁺和CD8⁺T淋巴细胞表面的PD-1和PD-L1表达水平均显著降低（P<0.05）;且观察组外周血CD4⁺和CD8⁺T淋巴细胞表面的PD-1和PD-L1表达水平均显著低于对照组（P<0.05）。结论 米诺环素治疗慢性牙周炎可有效改善患者各项牙周指标,改善牙周组织炎症反应抑制作用,在慢性牙周炎临床治疗中具有重要价值。     

NSCLC患者PD-L1表达对EGFR-TKIs疗效的影响

目的 分析非小细胞肺癌(NSCLC)患者程序性死亡分子1配体(PD-L1)表达对表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)疗效的影响.方法 回顾性分析首都医科大学大兴教学医院收治的94例EGFR敏感突变的Ⅳ期NSCLC患者的临床资料,所有患者均接受 EGFR-TKIs靶向治疗30 d以上.根据 PD-L1表...     

The PD-1 pathway as a therapeutic target to overcome immune escape mechanisms in cancer

Introduction: Immunotherapy is emerging as a powerful approach in cancer treatment. Preclinical data predicted the antineoplastic effects seen in clinical trials of programmed death-1 (PD-1) pathway inhibitors, as well as their observed toxicities. The results of early clinical trials are extraordinarily promising in several cancer types and have shaped the direction of ongoing and future studies.

Areas covered: This review describes the biological rationale for targeting the PD-1 pathway with monoclonal antibodies for the treatment of cancer as a context for examining the results of early clinical trials. It also surveys the landscape of ongoing clinical trials and discusses their anticipated strengths and limitations.

Expert opinion: PD-1 pathway inhibition represents a new frontier in cancer immunotherapy, which shows clear evidence of activity in various tumor types including NSCLC and melanoma. Ongoing and upcoming trials will examine optimal combinations of these agents, which should further define their role across tumor types. Current limitations include the absence of a reliable companion diagnostic to predict likely responders, as well as lack of data in early-stage cancer when treatment has the potential to increase cure rates.     

The PD-1/PD-L1 pathway: biological background and clinical relevance of an emerging treatment target in immunotherapy

Introduction: The co-inhibitory receptor programmed death 1 (PD-1) and its ligands are key regulators in a wide spectrum of immune responses and play a critical role in autoimmunity and self-tolerance as well as in cancer immunology. Emerging evidence suggests that cancer cells might use the PD-1/PD-ligand (PD-L) pathway to escape anti-tumor immunity. Based on this evidence, early phase human clinical trials targeting the PD-1/PD-L pathway are currently underway for multiple human cancers.

Areas covered: The role of the PD-1/PD-L pathway in autoimmune disease, viral infections as well as in malignant neoplasms is discussed and an overview of the existing therapeutics as well as the results of clinical trials targeting this pathway in cancer is given.

Expert opinion: The PD-1/PD-L pathway represents an important mechanism of immune evasion for malignant neoplasms. Early clinical trials indicate effectiveness of PD-1/PD-L pathway blockade in several solid cancers. However, greater insight into the exact mechanisms by which tumors are able to evade anti-tumor immunity is needed to increase clinical effectiveness, for example by combination blockade of diverse co-inhibitory receptors.     

PD-1/PD-L1单抗治疗晚期非小细胞肺癌引起免疫治疗相关性肺炎的网状Meta分析

目的:采用网状Meta分析(network meta-analysis,NMA),比较程序性细胞死亡蛋白-1及其配体(PD-1/PD-L1)单克隆抗体的不同治疗方案在晚期非小细胞肺癌(NSCLC)中导致免疫治疗相关性肺炎(IRP)的风险。方法:从PubMed、Embase和Cochrane Library数据库中检索PD-1/PD-L1单抗治疗晚期NSCLC的Ⅱ/Ⅲ期随机对照试验(RCT),检索时限为建库起至2021年6月1日。采用STATA 16.0和R软件4.1.0进行NMA。结果:共纳入30篇RCTs,包含18 425名患者。1~5级IRP的NMA结果显示,与化疗相比,PD-1单抗以及PD-L1单抗治疗均导致IRP的风险增高(P<0.05);PD-1单抗单药比PD-L1单抗单药及PD-L1单抗联合化疗发生IRP的风险增加(P<0.05);PD-1单抗+CTLA-4抑制剂发生IRP的风险高于PD-L1单抗、PD-1/PD-L1单抗+化疗(P<0.05);在3~5级IRP中,PD-1单抗、PD-1/PD-L1单抗+CTLA-4抑制剂发生IRP的风险高于PD-L1单抗+化疗(P<0.05);PD-1单抗+化疗发生重度IRP的风险低于PD-1单抗+CTLA-4抑制剂(P<0.05)。累积排序曲线下面积排序结果显示:发生1~5级和3~5级IRP可能性最高的均为PD-1单抗+CTLA-4抑制剂。结论:在晚期NSCLC的治疗中,PD-1/PD-L1单抗发生IRP的风险均高于化疗,PD-1单抗+CTLA-4抑制剂发生IRP的可能性最高,PD-1单抗发生IRP的风险高于PD-L1单抗。     

PD-1/PD-L1抑制剂联合其他免疫检查点抑制剂治疗三阴性乳腺癌的研究进展#br#

三阴性乳腺癌（TNBC）具有全身转移率高、对常规治疗不敏感以及容易产生耐药性等特点,导致患者的预后较差。随着对机体免疫系统抗肿瘤机制及TNBC免疫特点的不断探究,以程序性细胞死亡蛋白1（PD-1）和程序性死亡蛋白配体1（PD-L1）为代表的免疫检查点抑制剂为TNBC提供了新的治疗方案,但PD-1/PD-L1抑制剂单药治疗的效果不甚理想。本文就PD-1/PD-L1抑制剂联合其他具有不同机制的免疫检查点抑制剂在TNBC患者中的应用进行综述。     

Formononetin induces the mitochondrial apoptosis pathway in prostate cancer cells via downregulation of the IGF-1/IGF-1R signaling pathway

Context: Formononetin, an isoflavone, can inhibit the proliferation of cancer cells, including those of the prostate. However, its antitumor mechanism remains unclear.

Aim: To investigate whether the insulin-like growth factor 1 (IGF-1)/insulin-like growth factor 1 receptor (IGF-1?R) signaling pathway mediates the formononetin antitumor effect on prostate cancer cells.

Materials and methods: The viability of PC-3 cells was measured by MTT assay 48?h after formononetin treatment (25, 50 and 100?μM). Formononetin-induced cell apoptosis was measured by Hoechst 33258 staining and flow cytometry. Expression of Bax mRNA was detected by real-time PCR, and the expression levels of Bax and IGF-1?R proteins were detected by western blots.

Results: At concentrations >12.5?μM, formononetin significantly inhibited the proliferation of human prostate cancer cells. Formononetin increased Bax mRNA and protein expression levels and decreased the expression levels of pIGF-1?R protein in a dose-dependent manner.

Conclusion: High concentrations of formononetin-induced apoptosis in androgen-independent prostate cancer cells through inhibition of the IGF-1/IGF-1?R pathway.     

顺铂联合吉西他滨对肺癌PD-1/PD-L1途径的影响

目的 探讨PD-1/PD-L1途径在肺癌中表达情况及顺铂联合吉西他滨的干预效果.方法 将就诊于丽水市中心医院的80例肺癌患者随机分为对照组(A组)、顺铂治疗组(B组)、吉西他滨治疗组(C组)和顺铂联合吉西他滨治疗组(D)组,每组20例,收集各组患者手术切除的肿瘤组织,再以10例良性肺部疾病患者(E组)的手术切除组织作参照,采用Western blot法检测各组组织中PD-1、PD-L1的表达.选择10例同期来医院体检的健康人员(F组)外周血为空白对照,采用流式细胞仪检测B、C、D、F组治疗前后外周血中PD-L1+ CD68+巨噬细胞的表达,ELISA法检测B、C、D、F组治疗前后血清中IL-2、IL-4、IL-10的含量.结果 E组PD-1、PD-L1未见表达,A组PD-1、PD-L1的表达较E组上调(P＜0.01),B、C、D组较A组显著降低(P＜0.05),B、C组比较差异无统计学意义,D组与B、C组相比差异均有统计学意义(P＜0.05);B、C、D、F组均可检测到PD-L1+ CD68+巨噬细胞的表达,但B、C、D组中PD-L1+ CD68+巨噬细胞的比例显著高于F组(P＜0.01).B组和C组均可降低PD-L1+ CD68+巨噬细胞的比例(P＜0.05),而D组PD-L1+ CD68+巨噬细胞比例显著下调(P＜0.01).B、C、D组治疗前外周血血清中IL-2和IL-4含量较F组低(P＜0.01),IL-10含量较F组高(P＜0.01),治疗后B、C、D组外周血血清中IL-2和IL-4的含量较治疗前均有一定程度的上升(P＜0.05),IL-10含量有一定程度的降低(P＜0.05).结论 顺铂联合吉西他滨可下调肺癌组织中PD-1/PD-L1的阳性表达率及表达量,降低PD-L1+ CD68+巨噬细胞阳性表达,恢复PD-L1对IL-2、IL-4,IL-10所产生的影响,较单独用药效果明显.     

人程序化死亡因子5（PDCD5）在非小细胞肺癌细胞中的表达及相关性研究

目的：研究PDCD5在非小细胞肺癌中的表达及其相关性．探讨其在非小细胞肺癌发生、发展中的作用。方法：采用免疫组织化学方法,研究非小细胞肺癌组织中PDCD5的表达。结果：PDCD5阳性表达率随非小细胞肺癌组织学分级的降低和临床分期的上升而下降．染色强度也减弱。结论：PDCD5为非小细胞肺癌的负性调节因子,对抑制非小细胞肺癌的恶性转化和进展可能有重要的意义。     

Cdk5 knocking out mediated by CRISPR-Cas9 genome editing for PD-L1 attenuation and enhanced antitumor immunity

Blocking the programmed death-ligand 1 (PD-L1) on tumor cells with monoclonal antibody therapy has emerged as powerful weapon in cancer immunotherapy. However, only a minority of patients presented immune responses in clinical trials. To develop an alternative treatment method based on immune checkpoint blockade, we designed a novel and efficient CRISPR-Cas9 genome editing system delivered by cationic copolymer aPBAE to downregulate PD-L1 expression on tumor cells via specifically knocking out Cyclin-dependent kinase 5 (Cdk5) gene in vivo. The expression of PD-L1 on tumor cells was significantly attenuated by knocking out Cdk5, leading to effective tumor growth inhibition in murine melanoma and lung metastasis suppression in triple-negative breast cancer. Importantly, we demonstrated that aPBAE/Cas9-Cdk5 treatment elicited strong T cell-mediated immune responses in tumor microenvironment that the population of CD8⁺ T cells was significantly increased while regulatory T cells (Tregs) was decreased. It may be the first case to exhibit direct in vivo PD-L1 downregulation via CRISPR-Cas9 genome editing technology for cancer therapy. It will provide promising strategy for preclinical antitumor treatment through the combination of nanotechnology and genome engineering.     

FoxM1 mediated resistance to gefitinib in non-smallcell lung cancer cells

Aim:

Gefitinib is effective in only approximately 20% of patients with non-small-cell lung cancer (NSCLC), and the underlying mechanism remains unclear. FoxM1 is upregulated in NSCLC and associated with a poor prognosis in NSCLC patients. In this study, we examined the possible role of FoxM1 in gefitinib resistance and the related mechanisms.

Methods:

Gefitinib resistant human lung adenocarcinoma cell line SPC-A-1 and gefitinib-sensitive human lung mucoepidermoid carcinoma cell line NCI-H292 were used. mRNA and protein expression of FoxM1 and other factors were tested with quantitative RT PCR and Western blot analysis. RNA interference was performed to suppress FoxM1 expression in SPC-A-1 cells, and lentiviral infection was used to overexpress FoxM1 in H292 cells. MTT assay and flow cytometry were used to examine the proliferation and apoptosis of the cells.

Results:

Treatment of SPC-A-1 cells with gefitinib (1 and 10 μmol/L) upregulated the expression of FoxM1 in time- and concentration-dependent manners, while gefitinib (1 μmol/L) downregulated in H292 cells. In SPC-A-1 cells treated with gefitinib (1 μmol/L), the expression of several downstream targets of FoxM1, including survivin, cyclin B1, SKP2, PLK1, Aurora B kinase and CDC25B, were significantly upregulated. Overexpression of FoxM1 increased the resistance in H292 cells, while attenuated FoxM1 expression restored the sensitivity to gefitinib in SPC-A-1 cells by inhibiting proliferation and inducing apoptosis.

Conclusion:

The results suggest that FoxM1 plays an important role in the resistance of NSCLC cells to gefitinib in vitro. FoxM1 could be used as a therapeutic target to overcome the resistance to gefitinib.