174例早期肺腺癌患者EGFR基因突变检测和临床病理特征分析

目的　探讨早期肺腺癌组织中表皮生长因子受体（EGFR）基因突变和临床病理特征的关系。方法　选择2019年1月至2020年12月郑州人民医院收治的54例原位肺腺癌患者和120例微浸润肺腺癌患者作为研究对象,其中男性54例,女性120例,年龄25～79岁。利用扩增阻滞突变系统-聚合酶链反应（amplification refractory mutation system PCR,ARMS-PCR）方法检测以上组织中EGFR基因18、19、20、21外显子的突变情况,并对其与临床病理特征的相关性进行了回顾性研究。采用χ²检验。结果　受检的174例标本中,EGFR基因的总突变率为58.6%（102/174）,18、19、20、21外显子的突变分别占4.9%（5/102）、27.5%（28/102）、3.0%（3/102）、64.7%（66/102）。EGFR基因突变率的特点为：女性患者（64.2%,77/120）高于男性患者（46.3%,25/54）（P=0.027）;无吸烟史患者（64.7%,77/119）高于有吸烟史患者（45.5%,25/55）（P=0.017）;微浸润肺腺癌患者（65.0%,78/120）高于原位肺腺癌患者（44.4%,24/54）（P=0.011）;纯磨玻璃病灶（59.8%,67/112）和混杂磨玻璃病灶（63.5%,33/52）高于实性病灶（20.0%,2/10）（P=0.035）;肿物长径≥1 cm的（76.9%,60/78）明显高于肿物长径<1 cm的（43.8%,42/96）（P=0.001）;与年龄无明显相关性（P=0.804）。结论　在早期肺腺癌中,EGFR基因突变常见于女性、无吸烟史、微浸润肺腺癌、磨玻璃病灶、肿物长径≥1 cm的患者,以21号外显子L858R突变为主。EGFR基因突变在早期肺腺癌的发生发展过程中扮演重要角色。     

Luteolin is effective in the non-small cell lung cancer model with L858R/T790M EGF receptor mutation and erlotinib resistance

Background and Purpose

Non-small cell lung cancer (NSCLC) is one of the most commonly diagnosed malignancies in the world. Small-molecule inhibitors of the EGF receptor''s tyrosine kinase domain (TKIs), including gefitinib and erlotinib, have been widely used for treating NSCLC. Unfortunately, nearly all patients after initially experiencing a marked improvement while on these drugs, eventually progress to acquire resistance to TKIs. Because there is no effective therapeutic strategy to treat TKI-resistant NSCLC, we evaluated the effects of luteolin, a naturally occurring flavanoid, on T790M mutant NSCLC cells.

Experimental Approach

The effect of luteolin on the viability of NSCLC and normal cell lines was investigated using the Cell Counting Kit-8 (CCK-8) assay. Luteolin-induced apoptosis was assessed by bivariate FITC-annexin V/PI assay, and Western blots were used to measured apoptotic proteins. Co-immunoprecipitation was used to determine the effect of luteolin on the interaction between Hsp90 and mutant EGF receptors. The effect of luteolin on the Akt/mTOR pathway was studied using Western blotting analysis. Its anti-tumour efficacy in vivo was examined in a mouse xenograft model.

Key Results

Luteolin exerted significant anti-tumourigenic effects on the EGF receptor L858R/T790M mutation and erlotinib-resistant NSCLC both at the cellular and animal levels. Mechanistically, luteolin induced degradation of the EGF receptor by inhibiting the association of Hsp90 with the mutant EGF receptor, and, therefore, prevented PI3K/Akt/mTOR signalling, which resulted in NSCLC cell apoptosis.

Conclusion and Implications

Luteolin may be a potential candidate for NSCLC therapy, especially for treatment of patients with acquired erlotinib-resistant NSCLC.     

The role of EGFR tyrosine kinase inhibitors in the first-line treatment of advanced non small cell lung cancer patients harboring EGFR mutation

Lung cancer continues to be the leading cause of cancer death worldwide. Among lung cancers, 80% are classified as nonsmall- cell lung cancer (NSCLC) and are mostly diagnosed at an advanced stage (either locally advanced or metastatic disease). In the last years, the discovery of the pivotal role in tumorigenesis of the Epidermal Growth Factor Receptor (EGFR) has provided a new class of targeted therapeutic agents: the EGFR tyrosine kinase inhibitors (EGFR-TKIs). Since the first reports of an association between somatic mutations in EGFR exons 19 and 21 and response to EGFR-TKIs, treatment of advanced NSCLC has changed dramatically. Histologic profile, clinical characteristics, and mutational profile of lung carcinoma have all been reported as predictive factors of response to EGFR-TKIs and other targeted therapies. In advanced NSCLC patients harboring EGFR mutations, the use of EGFR TKIs in first-line treatment has provided an unusually large progression-free survival (PFS) benefit with a negligible toxicity when compared with cytotoxic chemotherapy in phase III randomized trials. Considering the findings regarding the excellent benefit and better safety profile of EGFR TKIs in EGFR mutation positive patients, these targeted therapeutic agents can be now considered as first-line treatment in this setting of patients. This review will discuss the new evidences in the role of EGFR-TKIs in the first-line treatment of advanced NSCLC and their implication in the current clinical decision-making.     

靶向联合化疗与常规化疗治疗表皮生长因子受体基因突变肺腺癌患者的临床效果

目的 探讨靶向联合化疗治疗表皮生长因子受体（EGFR）基因突变晚期肺腺癌患者的临床疗效及靶向联合化疗对EGFR基因不同突变位点患者的疗效差别。方法 选择安徽省胸科医院2016年1~12月收治确诊的64例EGFR基因检测阳性的Ⅲb/IV期肺腺癌患者,使用随机数字表方法分为靶向联合化疗组（33例）与常规化疗组（31例）,同时对靶向联合化疗组患者按照其基因突变位点不同分为3个亚组（19外显子突变组、21外显子突变组、20外显子突变组）。靶向联合化疗组患者采用EGFR受体酪氨酸抑制剂靶向治疗联合培美曲塞+卡铂/顺铂治疗,常规化疗组患者采用培美曲塞+卡铂/顺铂治疗。比较两治疗组患者的近期及远期疗效,并对靶向联合化疗组不同位点远期疗效进行数据分析。结果 靶向联合化疗组的中位无进展生存期高于常规化疗组,两组差异有统计学意义（P<0.05）,两组患者总体疗效和不良反应总发生率相似,差异无统计学意义（P>0.05）。靶向联合化疗组中,不同外显子突变的肺腺癌患者之间靶向联合化疗的生存期相似,差异无统计学意义（P>0.05）。结论 EGFR基因突变晚期肺腺癌患者接受靶向联合化疗能延长无进展生存时间,且不良反应未见增加。不同位点基因突变患者接受靶向联合化疗的临床效果无明显差别。     

非小细胞肺癌患者外周血中EGFR基因突变与埃克替尼治疗效果的相关性

目的 探讨非小细胞肺癌（NSCLC）患者外周血液中表皮生长因子受体（EGFR）基因突变与埃克替尼治疗效果的相关性。方法 选取洛阳市第三人民医院自2014年2月-2018年2月收治的101例NSCLC患者作为实验对象,采用RT-PCR技术检测外周血液中EGFR基因突变情况,依据测定结果分为基因突变组和野生型组,均采用埃克替尼治疗分析其治疗效果;选取同期做健康体检的患者50例作为对照组,比较NSCLC患者与健康人外周血液中EGFR突变差异。结果 50例健康人外周血液检测EGFR基因突变率为0,NSCLC患者中EGFR基因突变率为41.58%（42例）,EGFR野生型组患者31.68%（32例）,EGFR基因未突变者27例。基因突变组患者疾病控制率为85.71%,治疗有效率为64.29%;野生型组疾病控制率为59.38%,总有效率仅为12.5%,数据差异有统计学意义（P<0.05）。随访6个月内两组患者生存率差异无统计学意义,随访1、2年间EGFR基因突变组的生存率均远高于野生型组,数据差异有统计学意义（P<0.05）。结论 NSCLC患者外周血中EGFR基因突变患者行埃克替尼治疗效果更高,因此在晚期的NSCLC患者治疗中可以通过测定是否EGFR基因突变来指导靶向药物治疗。     

二代测序技术检测内蒙古地区晚期肺腺癌病人血浆循环肿瘤 DNA中表皮生长因子受体突变状态的应用价值

目的探讨二代测序技术检测血浆循环肿瘤 DNA(ctDNA)中表皮生长因子受体( EGFR)突变状态在内蒙古地区晚期肺腺癌病人中的应用价值。方法分析内蒙古自治区人民医院 2016年 11月至 2017年 12月符合条件的 50例晚期肺腺癌病人,采用突变阻滞扩增系统 PCR(ARMS-PCR)法检测组织样本、二代测序技术检测血浆 ctDNA中 EGFR突变情况,比较两种检测方法的一致性,并以组织检测作为金标准,评价二代测序技术的有效性。结果以组织检测结果为金标准,血浆 ctDNA中 EGFR突变检测的灵敏度为 96%,特异度为 68%,一致率为 84%。结论基于二代测序的血浆 EGFR突变检测与组织样本检测具有较好的一致性(Kappa=0.66,P=0.001)可以通过二代测序技术来反映晚期肺腺癌病人的 EGFR突变情况。     

埃克替尼治疗表皮生长因子受体突变型非小细胞肺癌患者49例

目的评价埃克替尼治疗表皮生长因子受体(EGFR)突变型非小细胞肺癌(NSCLC)患者的临床有效性及安全性。方法回顾性分析本院2011年12月—2013年12月年内住院的服用埃克替尼治疗的49例EGFR突变型NSCLC患者,以同期服用吉非替尼治疗的21例患者作为对照,比较两组的体能状态、生活质量评价及疗效和安全性评价。结果埃克替尼和吉非替尼组客观有效率分别为59%和57%,疾病控制率分别为84%和81%,两组相比无显著差异(P>0.05)。埃克替尼组在不良反应发生的类别、发生率及不良反应的分级等方面与吉非替尼组无显著差异(P>0.05)。结论埃克替尼治疗EGFR突变型NSCLC患者疗效确切、不良反应发生率较低,与吉非替尼相似。     

埃克替尼联合化疗治疗肺癌EGFR基因L833V和H835L罕见复合突变1例

非小细胞肺癌发生发展最常见的驱动因素是表皮生长因子受体（EGFR）突变,主要为19外显子缺失和第21外显子L858R突变,其他罕见基因突变发生率低,临床治疗方案尚未明确。本文报道1例携带EGFR基因H835L和L833V罕见突变的非小细胞肺癌患者,给予埃克替尼单药治疗3个月,疾病稳定（SD）,再予以埃克替尼联合化疗后病灶缩小,并维持SD近5个月。     

Less immune cell infiltration and worse prognosis after immunotherapy for patients with lung adenocarcinoma who harbored STK11 mutation

The STK11 mutation defined a special subtype for patients with lung adenocarcinoma. The cBioPortal data platform was applied to analyze STK11 mutation frequency and the relationship between STK11 mutation and immune prognostic markers. The TIMER database was used to analyze the relationship between STK11 mutation and immune cell infiltration. The survival difference for lung adenocarcinoma patients harbored STK11 mutation who received immunotherapy also used the cBioPortal database. The results showed that STK11 mutation co-occurrence more KRAS and KEAP1 mutation and fewer TP53 and EGFR mutation (all, P < 0.05); the patients harbored STK11 mutation had a lower expression of PDL1 (P = 0.002), higher TMB score (P = 0.002), higher proportion of males and smoking history; the patients harbored STK11 mutation had fewer immune cell infiltration including B cell (P < 0.01), CD8+ T cell (P < 0.001), CD4+ T cell (P < 0.001), Macrophage (P < 0.001), Neutrophil (P < 0.001) and Dendritic cell (P < 0.001). Importantly, we found the patients harbored STK11 mutation who received immune checkpoint inhibitors have worse overall survival (OS) with median survival only 6 months. In conclusion, our study demonstrated that STK11 mutation defined a special subtype for lung adenocarcinoma patients with different co-occurrence gene mutation, lower PDL1 expression, fewer immune cell infiltration and worse OS benefit from immune checkpoint inhibitors.     

First-line gefitinib for elderly patients with advanced NSCLC harboring EGFR mutations. A combined analysis of North-East Japan Study Group studies

Objective: To assess outcomes of elderly patients with advanced NSCLC harboring an EGFR mutation treated with gefitinib, as well as safety and impact on quality of life (QoL).

Methods: We performed a retrospective analysis of pooled data from one Phase III and two Phase II studies of 71 patients aged ≥ 70 years with a performance status of 0 – 2. The main outcome measures were progression-free survival (PFS), overall survival (OS) and response rate (RR), as well as incidence of adverse events and time to 9.1% deterioration in QoL.

Results: Median PFS (14.3 vs 5.7 months, p < 0.001) and overall RR (73.2 vs 26.5%, p < 0.001) in the gefitinib group were superior to those in the standard chemotherapy group, whereas median OS was not significantly different (30.8 vs 26.4 months, p = 0.42). Elevation of aspartate transaminase and/or alanine transaminase (18.3%) was the most common adverse event, and one treatment-related death (pneumonitis) occurred. Time to 9.1% deterioration in the QoL domains of pain and dyspnea, anxiety, and daily functioning was similar between the two age groups.

Conclusion: First-line gefitinib is efficacious with acceptable toxicity in relatively fit elderly patients with advanced NSCLC harboring an EGFR mutation.     

Brucine,an effective natural compound derived from nux-vomica,induces G1 phase arrest and apoptosis in LoVo cells

Brucine is an alkaloid from nux vomica, has been shown various pharmacological actions. To study the possible anti-cancer mechanisms on LoVo cells, effects of Brucine on cell viability, cell cycle and apoptosis were investigated. The results showed that Brucine revealed strong growth inhibitory effect on LoVo cells, and caused LoVo cell shrinkage and membrane blobbing, induced cellular and DNA morphological changes. Cell cycle and apoptosis analysis documented that Brucine could change cell cycle and induce cell apoptosis. Brucine-mediated cell cycle arrest in G1 phase was associated with a marked increase of protein levels of CCND1 and decrease in CCNB1, cyclin E and CDC2. In addition, Brucine dose-dependently caused LoVo cells apoptosis evidenced by Annexin V/PI staining Brucine-induced apoptosis was mediated via up-regulation of Bax and down-regulation of Bcl-2. Furthermore, proteins Erk1/2, p38 and Akt phosphorylation were down regulated by Brucine in a dose-dependent manner. In summary, this paper indicates Brucine is effective against LoVo cells proliferation, and promotes LoVo cells death via apoptosis. These results reveal functional interplay among a series of pathway that are deregulated in cancer and suggest that their simultaneous targeting by Brucine could result in efficacious inhibition on cancer cells.     

Erlotinib induces mitochondrial-mediated apoptosis in human H3255 non-small-cell lung cancer cells with epidermal growth factor receptorL858R mutation through mitochondrial oxidative phosphorylation-dependent activation of BAX and BAK

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor erlotinib shows potent antitumor activity in some non-small-cell lung cancer (NSCLC) cell lines and is approved by the Food and Drug Administration as second and third line treatment for NSCLC. However, the molecular mechanisms by which erlotinib induces apoptosis remain to be elucidated. Here, we investigated the effect of erlotinib on apoptotic signal pathways in H3255 cells with the EGFR(L858R) mutation. Erlotinib induces apoptosis associated with the activation of caspases in a dose- and time-dependent manner. Erlotinib did not alter the expression of apoptotic receptors FAS and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), although it induced caspase-8 activation and BID cleavage. In addition, cell death caused by erlotinib was not prevented by coincubation with FAS and TRAIL antagonists, ZB-4 monoclonal antibody and TRAIL/Fc recombinant, suggesting that erlotinib-induced apoptosis is not associated with receptor-mediated pathways. Erlotinib induces loss of mitochondrial membrane potential and release of cytochrome c and second mitochondria-derived activator of caspases/direct IAP binding protein with low pI from mitochondria. Furthermore, erlotinib causes BAX translocation to mitochondria, BAX and BAK conformational changes, and oligomerization. Erlotinib did not induce reactive oxygen species generation, and cotreatment with antioxidants did not alter erlotinib-induced activation of BAX and BAK and apoptosis. However, cotreatment with inhibitors of mitochondrial oxidative phosphorylation significantly blocked erlotinib-induced activation of BAX and BAK and cell death. Benzyloxycarbiny-VAD-fluoromethyl ketone had no effect on erlotinib-induced BAX and BAK activation but effectively prevented apoptosis. Overexpression of BCL-2 caused a significant attenuation of erlotinib-induced cell death, but no effect on BAX and BAK activation. Down-regulation of BAX and BAK gene expression with small interfering RNA led to an effective reduction of erlotinib-induced apoptosis. Our data indicate that activation of BAX and BAK plays a critical role in the initiation of erlotinib-induced apoptotic cascades.     

Darbepoetin alfa: an effective treatment with flexible and simplified dosing for anemia in patients with cancer

Anemia is common in patients with cancer or myelodysplastic syndrome. Erythropoietic therapy offers an effective way to manage anemia by increasing hemoglobin levels, decreasing transfusion requirements, and alleviating symptoms. We reviewed data showing the feasibility and effectiveness of treatment with the erythropoiesis-stimulating protein darbepoetin alfa at extended dosing intervals to treat anemia in patients with cancer receiving multicycle chemotherapy. We also explored the darbepoetin alfa's potential for treating anemia in patients with myelodysplastic syndrome. Data from clinical studies and drug therapy evaluations confirm that darbepoetin alfa administered weekly, every 2 weeks, and every 3 weeks corrects and maintains hemoglobin levels in patients with chemotherapy-induced anemia. In addition, the data demonstrate that both weight-based and fixed dosing with darbepoetin alfa are effective, and that early intervention to treat anemia has clinical benefits. Darbepoetin alfa also is an effective treatment for anemia in patients with cancer not receiving chemotherapy, at extended dosing intervals of at least 3 weeks. Extended dosing for anemia treatment can provide benefits for patients, caregivers, and clinicians because it reduces the number of clinic visits needed and permits synchronizing anemia treatment with chemotherapy cycles. Data from recent studies suggest that darbepoetin alfa is effective for treating anemia in patients with myelodysplastic syndrome; this potential use is being investigated further in ongoing studies. Thus, darbepoetin alfa is an attractive therapy option for patients with chemotherapy or cancer-induced anemia. It allows increased flexibility and simplified dosing and may offer some benefit in the treatment of anemia in patients with myelodysplastic syndrome.     

培美曲塞联合奈达铂二线治疗肺腺癌和肺大细胞癌的临床研究

目的观察培美曲塞联合奈达铂二线治疗晚期肺腺癌和肺大细胞癌的临床疗效。方法 32例一线治疗失败的晚期肺腺癌和肺大细胞癌患者采用培美曲塞联合奈达铂化疗4周期后,评价其疗效和不良反应。结果客观有效率为18.8%(6/32),疾病控制率为62.5%(20/32)。随访5～29个月,平均无疾病进展生存时间(PFS)为4.2个月,平均生存期(OS)为9.9个月,1年生存率为40.6%。ⅢB期客观有效率明显高于Ⅳ期,差异有统计学意义(P<0.05)。Ⅲ度以上不良反应包括白细胞减少2例(6.2%)、血红蛋白降低2例(6.2%)、血小板减少1例(3.1%)。结论培美曲塞联合奈达铂二线治疗肺腺癌和肺大细胞癌疗效确切,可提高生活质量,不良反应较轻,耐受性好,值得临床进一步推广研究。     

培美曲塞联合铂类药物治疗老年晚期肺腺癌的疗效与安全性分析

目的:本试验旨在评价70岁以上老年晚期肺腺癌患者一线应用培美曲塞联合铂类药物化疗的疗效和安全性。方法:试验纳入70岁以上ⅢB或Ⅳ期、既往未接受过化疗的晚期肺腺癌,接受培美曲塞联合顺铂或卡铂每3周1次治疗直到病情进展,对其疗效和安全性进行回顾性分析。结果:研究共入组22例老年患者,疗效评价完全缓解0例,部分缓解8例(36.4%),稳定12例(54.5%),进展2例(9.1%),疾病控制率90.9%。全组患者的中位PFS为5.9个月。18例(86.4%)出现1/2级的血液学或非血液学毒性,3例(13.6%)出现3/4级血液学毒性(中性粒细胞减少或贫血)和3级胃肠道反应。结论:培美曲塞联合铂类药物化疗方案对一般状态好的70岁以上老年晚期肺腺癌患者有较好的疗效和耐受性,可以作为其一线化疗的治疗选择。     

Addition of oral sildenafil to beraprost is a safe and effective therapeutic option for patients with pulmonary hypertension

Although sildenafil, an oral phosphodiesterase type-5 inhibitor, may offer benefits in the pharmacological management of pulmonary hypertension (PH), safety and effectiveness have not been studied during coadministration with beraprost, an oral prostacyclin analogue. To address this issue, we administered oral beraprost (40 microg) on day 1 and beraprost (40 microg) plus sildenafil (25 mg) on days 2 to 6 patients with moderate to severe PH. Although sildenafil plus beraprost produced transient flushing in 2 of 6 patients, systemic hemodynamics and arterial and venous gas analyses were similar in comparisons between the 2 treatment groups. In contrast, sildenafil plus beraprost therapy resulted in a 2.2-fold greater reduction in mean pulmonary arterial pressure and a 1.6-fold greater reduction in pulmonary vascular resistance compared with beraprost alone, and reductions in these parameters persisted longer with combination therapy than with beraprost alone. Addition of oral sildenafil to beraprost appears to represent a safe and effective therapeutic option, at least in the acute phase, for patients with pulmonary hypertension.     

达可替尼在真实世界中治疗EGFR突变阳性晚期非小细胞肺癌患者的临床疗效和安全性

目的 探讨达可替尼在表皮生长因子受体(EGFR)突变阳性晚期非小细胞肺癌(NSCLC)患者的治疗效果及安全性.方法 回顾性分析50例EGFR突变阳性晚期NSCLC患者的临床资料,其中的24例(A组)给予达可替尼治疗,26例(B组)给予第一代表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)治疗.比较两组患者临床结局和...