Progress of Immunotherapy in EGFR-mutated Advanced Non-small Cell Lung Cancer / 中国肺癌杂志

Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) are currently the first-line standard of care for patients with non-small cell lung cancer (NSCLC) that harbor EGFR mutations. Nevertheless, resistance to EGFR-TKIs is inevitable. In recent years, although immune checkpoint inhibitors (ICIs) have significantly shifted the treatment paradigm in advanced NSCLC without driver mutation, clinical benefits of these agents are limited in patients with EGFR-mutated NSCLC. Compared with wild-type tumors, tumors with EGFR mutations show more heterogeneity in the expression level of programmed cell death ligand 1 (PD-L1), tumor mutational burden (TMB), and other tumor microenvironment (TME) characteristics. Whether ICIs are suitable for NSCLC patients with EGFR mutations is still worth exploring. In this review, we summarized the clinical data with regard to the efficacy of ICIs in patients with EGFR-mutated NSCLC and deciphered the unique TME in EGFR-mutated NSCLC.
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Mechanism of Stemona tuberosa Alkaloids in Inhibiting Proliferation and Inducing Apoptosis of Non-small Cell Lung Cancer NCI-H460 Cells / 中国实验方剂学杂志

ObjectiveTo study the effect and underlying mechanism of Stemona tuberosa alkaloids on the proliferation and apoptosis of human non-small cell lung cancer NCI-H460 cells. MethodNon-small cell lung cancer NCI-H460 cells were divided into a blank group and S. tuberosa alkaloids groups （50， 100， 150， 200， and 250 mg·L-1）. The effect of S. tuberosa alkaloids on the proliferation of human NCI-H460 cells was observed by thiazolyl blue tetrazolium bromide （MTT） assay and colony formation assay. Cell apoptosis was observed by Hoechst 33258 staining and flow cytometry. Real-time fluorescence-based polymerase chain reaction （Real-time PCR） was used to detect the effect of S. tuberosa alkaloids on the mRNA expression of cysteinyl aspartate-specific protease 3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and epidermal growth factor receptor （EGFR）. The protein expression levels of Caspase-3， Bax， Bcl-2， protein kinase B （Akt）， phosphorylated （p-）Akt， EGFR， c-Jun N-terminal kinase （JNK）， p-JNK， p38 mitogen-activated protein kinase （p38 MAPK）， and p-p38 MAPK were measured by Western blot. ResultCompared with the blank group， the S. tuberosa alkaloids groups showed increased inhibition rate on cell proliferation （P<0.01）， reduced number of cell clones formed and the rate of cell clonal formation （P<0.05， P<0.01）， and increased karyopyknosis， cytoplasmic aggregation， and cell apoptosis rate （P<0.01）. The S. tuberosa alkaloids groups at 100， 150， 200， and 250 mg·L-1 showed increased Caspase-3 mRNA expression （P<0.05）， decreased EGFR mRNA expression （P<0.05， P<0.01）， up-regulated protein expression of Caspase-3 and p-JNK （P<0.01）， and down-regulated protein expression of EGFR and p-Akt （P<0.05， P<0.01）. Additionally， compared with the blank group， the S. tuberosa alkaloids groups showed increased expression of Bax mRNA （P<0.01）， decreased expression of Bcl-2 mRNA （P<0.01）， up-regulated protein expression of Bax and p-p38 MAPK （P<0.01）， and down-regulated protein expression of Bcl-2 （P<0.01）. ConclusionsS. tuberosa alkaloids can inhibit proliferation and induce apoptosis of human non-small cell lung cancer NCI-H460 cells， and the mechanism may be related to the inhibition of EGFR protein expression and phosphorylation of Akt protein， as well as the activation of the JNK/p38 MAPK signaling pathway.

Anti-tumor Activity of Chinese Medicine Based on EGFR Signaling Pathway： A Review / 中国实验方剂学杂志

In China， malignant tumors have become the main cause of death. In the past half century， the incidence and mortality of malignant tumors have been on the rise， posing a threat to health of patients， and the burden of cancer has been increasing. At the moment， malignant tumors are mainly treated by surgery， radiotherapy， and cytotoxic drugs， which， however， have limitations and induce great adverse reactions. As biological technology and the research on tumor microenvironment， immunology， cell biology， and molecular biology advance， high-efficiency low-toxicity targeted therapy has attracted wide attention in the treatment of tumors. Epidermal growth factor receptor （EGFR） plays an important role in many cellular processes such as cell proliferation， survival， differentiation， migration， inflammation， and stromal homeostasis. EGFR promotes tumor growth， proliferation， and metastasis in a variety of ways. Chinese medicine has unique efficacy in the comprehensive treatment of malignant tumors. Through multiple components， multiple targets， and multiple pathways， it enhances the efficacy， reduces toxicity， prolongs life， and improves life quality in the treatment of tumors. Many Chinese medicines and their active components exert anti-tumor effect by inhibiting the EGFR signal transduction pathway. Through targeted inhibition of EGFR， Chinese medicine can promote the apoptosis and autophagy of tumor cells， suppress the proliferation and metastasis of tumor cells， and delay the progression of tumors. Thus， EGFR is a potential target for suppressing tumor. This paper summarizes the relationship between EGFR signal transduction pathway and tumor occurrence and development and analyzes the anti-tumor action mode and possible mechanisms of Chinese medicine and the active components by regulating EGFR signaling pathway， which is expected to provide a reference for clinical practice.

USH2A mutation and specific driver mutation subtypes are associated with clinical efficacy of immune checkpoint inhibitors in lung cancer / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

This study aimed to identify subtypes of genomic variants associated with the efficacy of immune checkpoint inhibitors (ICIs) by conducting systematic literature search in electronic databases up to May 31, 2021. The main outcomes including overall survival (OS), progression-free survival (PFS), objective response rate (ORR), and durable clinical benefit (DCB) were correlated with tumor genomic features. A total of 1546 lung cancer patients with available genomic variation data were included from 14 studies. The Kirsten rat sarcoma viral oncogene homolog G12C (KRASG12C) mutation combined with tumor protein P53 (TP53) mutation revealed the promising efficacy of ICI therapy in these patients. Furthermore, patients with epidermal growth factor receptor (EGFR) classical activating mutations (including EGFRL858R and EGFRΔ19) exhibited worse outcomes to ICIs in OS (adjusted hazard ratio (HR), 1.40; 95% confidence interval (CI), 1.01‍‒‍1.95; P=0.0411) and PFS (adjusted HR, 1.98; 95% CI, 1.49‍‒‍2.63; P<0.0001), while classical activating mutations with EGFRT790M showed no difference compared to classical activating mutations without EGFRT790M in OS (adjusted HR, 0.96; 95% CI, 0.48‍‒‍1.94; P=0.9157) or PFS (adjusted HR, 0.72; 95% CI, 0.39‍‒‍1.35; P=0.3050). Of note, for patients harboring the Usher syndrome type-2A(USH2A) missense mutation, correspondingly better outcomes were observed in OS (adjusted HR, 0.52; 95% CI, 0.32‍‒‍0.82; P=0.0077), PFS (adjusted HR, 0.51; 95% CI, 0.38‍‒‍0.69; P<0.0001), DCB (adjusted odds ratio (OR), 4.74; 95% CI, 2.75‍‒‍8.17; P<0.0001), and ORR (adjusted OR, 3.45; 95% CI, 1.88‍‒‍6.33; P<0.0001). Our findings indicated that, USH2A missense mutations and the KRASG12Cmutation combined with TP53 mutation were associated with better efficacy and survival outcomes, but EGFR classical mutations irrespective of combination with EGFRT790M showed the opposite role in the ICI therapy among lung cancer patients. Our findings might guide the selection of precise targets for effective immunotherapy in the clinic.

CUDC-101 as a dual-target inhibitor of EGFR and HDAC enhances the anti-myeloma effects of bortezomib by regulating G2/M cell cycle arrest / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

CUDC-101, an effective and multi-target inhibitor of epidermal growth factor receptor (EGFR), histone deacetylase (HDAC), and human epidermal growth factor receptor 2 (HER2), has been reported to inhibit many kinds of cancers, such as acute promyelocytic leukemia and non-Hodgkin's lymphoma. However, no studies have yet investigated whether CUDC-101 is effective against myeloma. Herein, we proved that CUDC-101 effectively inhibits the proliferation of multiple myeloma (MM) cell lines and induces cell apoptosis in a time- and dose-dependent manner. Moreover, CUDC-101 markedly blocked the signaling pathway of EGFR/phosphoinositide-3-kinase (PI3K) and HDAC, and regulated the cell cycle G2/M arrest. Moreover, we revealed through in vivo experiment that CUDC-101 is a potent anti-myeloma drug. Bortezomib is one of the important drugs in MM treatment, and we investigated whether CUDC-101 has a synergistic or additive effect with bortezomib. The results showed that this drug combination had a synergistic anti-myeloma effect by inducing G2/M phase blockade. Collectively, our findings revealed that CUDC-101 could act on its own or in conjunction with bortezomib, which provides insights into exploring new strategies for MM treatment.

Concomitant TP53 mutation in early-stage resected EGFR-mutated non-small cell lung cancer: a narrative approach in a genetically admixed Brazilian cohort

TP53 mutations are frequent in non-small cell lung cancer (NSCLC) and have been associated with poor outcome. The prognostic and predictive relevance of EGFR/TP53 co-mutations in NSCLC is controversial. We analyzed lung tissue specimens from 70 patients with NSCLC using next-generation sequencing to determine EGFR and TP53 status and the association between these status with baseline patient and tumor characteristics, adjuvant treatments, relapse, and progression-free (PFS) and overall survival (OS) after surgical resection. We found the EGFR mutation in 32.9% of patients (20% classical mutations and 12.9% uncommon mutations). TP53 missense mutations occurred in 25.7% and TP53/EGFR co-mutations occurred in 43.5% of patients. Stage after surgical resection was significantly associated with OS (P=0.028). We identified an association between progression-free survival and poor outcome in patients with distant metastases (P=0.007). We found a marginally significant difference in OS between genders (P=0.057) and between mutant and wild type TP53 (P=0.079). In univariate analysis, distant metastases (P=0.027), pathological stage (IIIA-IIIB vs I-II; P=0.028), and TP53 status (borderline significance between wild type and mutant; P=0.079) influenced OS. In multivariable analysis, a significant model for high risk of death and poor OS (P=0.029) selected patients in stage IIIA-IIIB, with relapse and distant metastases, non-responsive to platin-based chemotherapy and erlotinib, with tumors harboring EGFR uncommon mutations, with TP53 mutant, and with EGFR/TP53 co-mutations. Our study suggested that TP53 mutation tends to confer poor survival and a potentially negative predictive effect associated with a non-response to platinum-based chemotherapy and erlotinib in early-stage resected EGFR-mutated NSCLC.

Effect of Guiqi Baizhu Prescription Combined with Oxaliplatin on Expression of EGFR and VEGFR2 and Angiogenesis in Gastric Cancer-bearing Mice / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Guiqi Baizhu prescription （GQBZ） combined with oxaliplatin on the expression of epidermal growth factor receptor （EGFR） and vascular endothelial growth factor receptor-2 （VEGFR2） and angiogenesis in gastric cancer-bearing mice. MethodThe tumor-bearing model of gastric cancer was induced in Kunming mice. The mice were randomly divided into blank group， model group， oxaliplatin group （10 mg·kg-1）， and high- （17.68 g·kg-1）， medium- （8.84 g·kg-1）， and low-dose （4.42 g·kg-1） combination groups （GQBZ combined with oxaliplatin）. After the last administration， the transplanted tumor was collected and the tumor inhibition rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of tumor tissues. Enzyme-linked immunosorbent assay （ELISA） was used to detect the serum content of epidermal growth factor （EGF）， interleukin-8 （IL-8）， and vascular endothelial growth factor （VEGF）. Western blot and immunohistochemistry （IHC） were used to detect the expression of EGFR， phosphorylated EGFR （p-EGFR）， VEGFR2， phosphorylated VEGFR2 （p-VEGFR2）， and platelet-endothelial cell adhesion molecule （CD31）. Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of EGFR and VEGFR2. ResultThe tumor weight in the drug intervention groups was significantly lower than that in the model group （P<0.01）. Compared with the oxaliplatin group， the high- and medium-dose combination groups showed reduced tumor weight （P<0.05， P<0.01）. The tumor cells in the model groups were high in cell density and regular in shape， and no clear tissue necrosis was seen. The tumor cell density in the drug intervention groups was reduced， and clear tissue necrosis and large-scale inflammatory cells were visible. Compared with the blank group， the model group and the drug intervention groups showed increased serum levels of EGF， VEGF， and IL-8 （P<0.05， P<0.01）. Compared with the model group， the drug intervention groups showed decreased serum levels of EGF， VEGF， and IL-8 （P<0.01）， reduced protein expression of EGFR， p-EGFR， VEGFR2， p-VEGFR2， and CD31， and declining mRNA expression of EGFR and VEGFR （P<0.01）. Compared with the oxaliplatin group， the high- and medium-dose combination groups showed decreased serum levels of EGF， VEGF， and IL-8 （P<0.05， P<0.01）， reduced protein expression of EGFR， p-EGFR， VEGFR2， p-VEGFR2， and CD31， and dwindled mRNA expression of EGFR and VEGFR2 （P<0.05， P<0.01）. The low-dose combination group showed decreased serum levels of EGF， VEGF， and IL-8， reduced protein expression of EGFR， p-EGFR， VEGFR2， p-VEGFR2， and CD31， and dwindled mRNA expression of EGFR and VEGFR2， but the difference was not statistically significant. ConclusionGQBZ combined with oxaliplatin can inhibit the growth and angiogenesis of tumor tissues in gastric cancer-bearing mice by affecting the expression of EGFR and VEGFR2.

Effect of Maxing Shigantang on Airway Injury and EGFR Expression in Asthma Model Rats / 中国实验方剂学杂志

ObjectiveTo explore the optimal formula of Maxing Shigantang in regulating epidermal growth factor receptor（EGFR）expression and alleviating airway injury in asthmatic rats and to reveal the underlying mechanism. MethodSD male rats were randomly divided into normal group, model group, dexamethasone group (5×10-4 g·kg-1) and Maxing Shigantang 1∶0.5, 1∶1, 1∶2 groups (group A, B, C, 10 g·kg-1), with 8 rats in each group. The other groups except the normal group received nebulization of 2% acetylcholine chloride and 0.4% histamine phosphate for the modeling of asthma. One hour before modeling， the normal group and the model group were given the same amount of normal saline， and the other groups were given the same amount of corresponding drugs， once a day for 7 days. On the 7th day， the model was established and the incubation period of asthma was recorded. The rats were then immediately anesthetized， and arterial blood and tracheal tissue were collected. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the levels of interleukin-2 （IL-2）， interleukin-4 （IL-4）， and tumor necrosis factor-α （TNF-α） in serum. Pathological sections were prepared for the observation of the pathological changes of tracheal tissues and the ultrastructure of epithelial cells in each group. Terminal-deoxynucleotidyl transferase-mediated nick-end labeling （TUNEL） was adopted to detect epithelial cell apoptosis， and in situ hybridization and Western blot were employed to determine the mRNA and protein levels of epidermal growth factor receptor （EGFR）， respectively. ResultCompared with the model group， groups A， B and C prolonged the incubation period of asthma （P<0.05，P<0.01）. Compared with the control group， the model group showed declined IL-2 level （P<0.01）， risen IL-4 and TNF-α levels （P<0.05，P<0.01）， increased airway pathology score， collagen volume fraction， and airway epithelial cell apoptosis index （P<0.01）， and up-regulated mRNA and protein levels of EGFR in trachea tissue （P<0.01）. Compared with the model group， group A showed increased IL-2 level （P<0.05） and declined IL-4 （P<0.05，P<0.01） level， and group B showed declined IL-4 level （P<0.05）. The level of TNF-α in groups A， B， and C declined compared with that in the model group （P<0.01）. Maxing Shigantang repaired the tracheal tissue to different degrees （P<0.05）. Among the three groups， group A inhibited tracheal fibrosis （P<0.05） and had the most significant effect of repairing the ultrastructural changes of airway epithelial cells. Groups A， B and C all inhibited the apoptosis of airway epithelial cells （P<0.05）. All the three groups inhibited the up-regulation of EGFR mRNA level （P<0.05，P<0.01）， and groups B and C inhibited the up-regulation of EGFR protein level （P<0.05，P<0.01）. ConclusionMaxing Shigantang can inhibit the abnormal changes of airway epithelial structure， alleviate airway injury， and can down-regulate the expression of EGFR in the tracheal tissue of asthma model rats. In this study， the optimal compatibility of Maxing Shigantang to repair airway epithelial injury in asthmatic rats was group A， with the Ephedrae Herba-Armeniacae Semen Amarum-Glycyrrhizae Radix et Rhizoma-Gypsum Fibrosum ratio of 1∶0.5∶4∶1.

Preparation of anti EGFR scFv::FTH1/FTH1 nanoparticles for asthma treatment / 生物工程学报

Asthma is a common respiratory disease that affects 300 million of people worldwide, posing a serious health risk and medical burden. Development of new anti-asthmatic drugs and alternative treatment regimens is therefore encouraged. Recent studies have shown that Epidermal Growth Factor Receptor (EGFR) is involved in asthma development. In order to construct nanoparticles targeting EGFR for asthma treatment, a single chain antibody fragment (scFv) against EGFR was genetically engineered and modified at the N-terminal end of the human ferritin H-chain (FTH1) to construct Anti EGFR scFv::FTH1/FTH1 nanoparticles. Transmission electron microscopy showed that the nanoparticles were self-assembled into hollow cage-like structures with the particle size of about 12 nm. Semi-quantitative analysis of the purified nanoparticles by SDS-PAGE revealed the mass ratio of FTH1 to Anti EGFR scFv::FTH1 was 7:3. In House Dust Mite (HDM) driven models, Anti EGFR scFv::FTH1/FTH1 nanoparticles efficiently attenuated several key features of asthma, including goblet cell hyperplasia, mucous metaplasia and subepithelial fibrosis, showing the potential of using ferritin based nanoparticle for asthma treatment.

Genomic mutations in patients with advanced non-small cell lung cancer in Yunan and its clinical significance / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the lung cancer-associated driver gene mutations in peripheral blood of patients with advanced non-small cell lung cancer (NSCLC) in Yunnan area, and to explore their association with clinical pathological features. Methods: Peripheral blood of 304 patients with stage Ⅳ NSCLC were collected from Molecular Diagnostic Center of Yunnan Cancer Hospital during January 2019 to December 2019. Next generation sequencing (NGS) technique was used to detect the mutation of NSCLC related driver genes, chi-square test was used to analyze the relationship between the major mutant genes and the clinicopathological features of patients, and Logistic regression was used to analyze the independent risk factors. Results: In the peripheral blood of 304 patients with stage Ⅳ NSCLC, there were 120 (39.47%) cases with EGFR mutations, 12 (3.95%) cases with ALK fusion, 36 (11.84%) case with other mutations such as KRAS, BRAF and RET. The main EGFR mutations were 19del and L858R (69.17%). The mutation rate of EGFR was higher in female, young, non-smoking, non-chemotherapy and lung adenocarcinoma patients (49.26% vs 31.55%, 45.39% vs 33.56%, 45.92% vs 27.78%, 45.07% vs 26.37%, 42.39% vs 10.71%, all P<0.05). Multivariate analysis showed that female, no history of chemotherapy and lung adenocarcinoma were independent risk factors for EGFR mutations (all P<0.05). Conclusion: Using NGS technology to detect the driver genes in peripheral blood of patients with advanced NSCLC in Yunnan area showed that the mutation rate of EGFR was higher in women and lung adenocarcinoma patients without chemotherapy history.

lncRNA DGCR5 promotes the malignant biological behaviors of esophageal squamous cell carcinoma TE1 cells by up-regulating EGFR expression / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of long non-coding RNA DiGeorge syndrome critical region gene 5 (DGCR5) on proliferation, invasion and migration of esophageal squamous cell carcinoma (ESCC) TE1 cells and its mechanism. Methods: qPCR was used to detect the expression level of DGCR5 in ESCC cell lines (TE1, Yes-2, KYSE150 and Eca9706). TE1 cells were transfected with siRNA-DGCR5(si-DGCR5) and negative control (si-NC) plasmids, respectively. CCK-8, Wound healing and Transwell assay were used to detect the proliferation, migration and invasion of TE1 cells before and after DGCR5 knockdown. The relationship between DGCR5 expression and epidermal growth factor receptor (EGFR) in ESCC tissues was analyzed by GEPIA database. The mRNA and protein expressions of EGFR in ESCC cell line were examined by qPCR and Western blotting (WB). WB was further used to detect the expression of EGFR protein in TE1 cells before and after DGCR5 knockdown. Results: lncRNA DGCR5 was highly expressed in ESCC cell lines (all P<0.01). qPCR confirmed that the expression of DGCR5 in TE1 cells of si-DGCR5 group was significantly lower than that of si-NC group (P<0.01). The proliferation, migration and invasion ability of TE1 cells in si-DGCR5 group were significantly lower than those in si-NC group (all P<0.01). GEPIA database showed that the expression of DGCR5 was positively correlated with EGFR in ESCC tissues (P<0.01). WB showed that the protein level of EGFR in TE1 cells of si-DGCR5 group decreased significantly (P<0.01). Conclusion: lncRNA DGCR5 is highly expressed in ESCC cells, and promotes the proliferation, invasion and migration of TE1 cells possibly by up-regulating EGFR expression.

Charaoteristics and clinical significance of immune microenvironment in nonsmall cell lung cancer tissues / 中国肿瘤生物治疗杂志

@# Objective: To investigate the characteristics and clinical significance of the immunomicroenvironment typing based on the expression of programmed death-ligand 1 (PD-L1) and the infiltration of CD8+ T cells in the stroma in patients with non-small cell lung cancer (NSCLC). Methods: Paraffin tissue specimens and relevant clinicopathological data of 74 NSCLC patients admitted to our hospital from January 2016 to July 2018 were collected.All patients received EGFR gene test, and none received radiotherapy, chemotherapy or targeted therapy. Immunohistochemistry was used to detect the expression of PD-L1 in tissues and the infiltration of CD8+T cells in interstitium, and the relationship between PD-L1, CD8+T cells, and the immune microenvironment typing based on both, and the pathological parameters and the survival of patients was analyzed. Results: PD-L1 expression in the primary tumor of NSCLC patients showed statistical differences in gender, pathological type, smoking history, EFGR gene mutation status ( P <0.05). The infiltration of CD8+ T lymphocytes in tumor microenvironment showed statistically significant differences in different TNM stage and lymph node metastasis ( P <0.05), PD-L1 expression was significantly correlated with EGFR mutation ( P =0.000), while CD8+T lymphocyte infiltration was not correlated with EGFR mutation ( P =0.605). The immunomicroenvironment of EGFR wild-type patients was mainly (CD8+ PD-L1+) (type I), and the mutants were mainly (CD8-PD-L1-) (type II) and (CD8+PD-L1-) (type IV). The distribution of immune microenvironmental typing in each group with different EGFR mutation, smoking history and pathological differentiation degree was significantly different ( P <0.05) and significantly correlated with EGFR mutation ( P <0.05). Follow-up showed that the patients with disease free survival, recurrence and metastasis and death were the most in type I, type II and type I, respectively. Conclusions: In this study, the distribution of tumor immunomicroenvironmental typing in NSCLC patients was mainly the highest in type I and the lowest in type Ⅲ, which was related to EGFR mutation, smoking history and pathological differentiation. Patients with EGFR mutations were mainly of type Ⅱand type Ⅳ, and were associated with low expression of PD-L1. ··

Detection of EGFR gene mutation in periphral blood of NSCLC patients in Yunnan by Super-ARMS and its clinical significance / 中国肿瘤生物治疗杂志

@#Objective: To detect the mutation of epidermal growth factor receptor (EGFR) gene in peripheral blood of non-small cell lung cancer (NSCLC) patients in Yunnan area with Super-ARMS, and to explore its correlation with clinicopathological characteristics. Methods: A total of 222 blood samples from patients with NSCLC were collected between January 2017 to December 2018 in the Molecular Diagnostic Center of Yunnan Cancer Hospital. The EGFR gene mutation in peripheral blood samples was detected by SuperARMS, and the relationship between EGFR gene mutation and clinicopathological features was analyzed. Meanwhile, the independent risk factors influencing EFGR mutation were also analyzed. Results: In the peripheral blood of 222 NSCLC patients, there were 81 cases (36.5%) with EGFR gene mutation. Among them, exon 19 deletion and L858R gene point mutation were the most common (75.3% of total mutation); female patients had a higher mutation rate than male patients (45.9% vs 27.0%); patients <60 years old had a higher incidence of mutation than patients≥60 years old (43.2% vs 28.8%) (P<0.05 or P<0.01); moreover, patients with no history of smoking, no history of radical surgery, adenocarcinoma, advanced stage and no history of chemotherapy had higher incidence of EGFR mutation (43.9% vs 21.6%, 39.2% vs 21.2%, 43.9% vs 4.8%, 39.7% vs 23.3% and 44.0% vs 23.5%) (P<0.05 or P<0.01). Multivariate logistic analysis showed that young, no smoking history, adenocarcinoma and no surgical history were independent risk factors for EGFR gene mutation (all P<0.01). Conclusion: In the peripheral blood of patients with NSCLC in Yunnan, the mutation rate of EGFR gene is higher in patients with age<60 years old, adenocarcinoma and non-smoking. Super-ARMS method is more sensitive in the detection of EGFR mutation in peripheral blood of lung cancer patients.

Advances in the study of microRNA-7 involved in the injury and repair of intestinal epithelial barrier via EGFR / 国际儿科学杂志

MicroRNAs (miRNAs) are non-coding single-stranded small RNAs composed of 19 ～25 nucleotides involved in various life processes such as cell proliferation,differentiation,apoptosis,signal transduction and immune regulation at the post-transcriptional level.So it has become one of the research hotspots in recent years.Among them,MiR-7 is considered to be an important regulator of epithelial cell damage and repair.As the largest epithelium of the human body,the intestinal macosal epithelium is the link between the human body and the outside world,and plays an important role in the homeostasis of the internal environment.Numerous studies have shown that MiR-7 plays an irreplaceable role in intestinal epithelial barrier damage and repair through the epidermal growth factor receptor (EGFR)-related signal pathway.This paper reviews the research progress.

Advanced Research on Non-small Cell Lung Cancer with De Novo T790M Mutation / 中国肺癌杂志

With the development of sequencing technology, the detection rate of de novo T790M mutation is increasing. The emergence of the third generation of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) provide treatment opportunities. Secondary T790M mutation is often emphasized in clinic, but de novo T790M mutation is neglected. This review found that the incidence of de novo T790M mutation fluctuated greatly, which was mainly affected by sequencing techniques. The de novo T790M mutation is mainly low in mutation abundance, easy to combine with other gene changes, a poor predictor and prognostic factor and the efficacy of the first and second generation EGFR-TKIs is limited. The therapeutic value of osimertinib needs to be studied.
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Advances in the treatment of patients with stageⅡ-ⅢA non-small cell lung cancer and the EGFR mutation / 中国肿瘤临床

The incidence of non-small cell lung cancer (NSCLC) is the highest among malignant tumors. StagesⅡtoⅢA patients are potentially treatable. Perioperative chemotherapy is the standard treatment for NSCLC; however, in patients with the EGFR mutation, neoadjuvant or adjuvant targeted therapy can improve disease-free survival (DFS). Despite this, the survival benefit is not clear. Postop-erative adjuvant radiotherapy can enhance survival in patients with stageⅢA (N2) NSCLC. Durvalumab maintenance therapy, after concurrent chemoradiotherapy, has become the new standard for patients with locally advanced and unresectable NSCLC. Immuno-therapy requires additional investigation in patients with driver gene mutations, as does the combination of immunotherapy plus che-motherapy. Notably, anti-angiogenesis therapy has failed as a postoperative adjuvant therapy.

Ergosta-7,22-diene-2β,3α,9α-triol (EGDT) from Ganoderma lucidum inhibits nasopharyngeal carcinoma cells by blocking EGFR signaling pathway / 中草药·英文版

Objective: To investigate the efficacy and mechanism of EGDT against NPC cell lines. Methods: MTT assay was used to assess cell proliferation inhibition of EGDT. The apoptotic induction and cell cycle arrest were detected by flow cytometry. Western blot was adopted to detect the protein levels. Quantitative Real-time PCR was used to determine the mRNA expressions. The NPC xenografts were established to evaluate the tumor growth inhibition of EGDT. Immunohistochemistry was applied to analyze the EGFR expression in the tumor tissues. Results: EGDT showed proliferation inhibition on the NPC cell, induced G0/G1 phase arrest and cell apoptosis in vitro. EGDT decreased the protein and mRNA levels of EGFR and its downstream RAF/MEK/ERK and PI3K/AKT pathways in time- and dose-dependent manner. Furthermore, EGDT also showed a sound antitumor activity in NPC xenograft in vivo. Conclusion: The treatment of EGDT displays EGFR and its mediated downstream signaling pathway blockade through decreasing the protein and mRNA levels, suggesting a promising strategy in treating human NPC.

Advances in Double Mutations of EGFR and ALK Gene in Non-small Cell Lung Cancer / 中国肺癌杂志

Molecular target therapy is one of the most popular field of non-small cell lung cancer (NSCLC) treatmnet. Epidermal growth factor receptor (EGFR) mutation and anaplastic lymphoma kinase (ALK) rearragement are the most important two oncogenic drivers in NSCLC, early studies suggested that EGFR mutations and ALK rearrangements are mutually exclusive, but isolated cases or small sample research with concomitant EGFR and ALK alterations have been constantly reported. The co-occurrence of EGFR mutations and anaplastic lymphoma kinase (ALK) rearrangements constitutes a rare molecular, the frequency of EGFR/ALK co-alterations was about 1%, however, little has been known about clinicopathologic feature and treatment. This review summarized published case report, EGFR and ALK alterations are common in female, Asian origin, never smoker, IV stage, and denocarcinomas. First-line treatment can choose EGFR or ALK tyrosine kinase inhibitors (TKIs). However, studies about the origin and resistance mechanism in EGFR/ALK co-alterations are little, require more experimental and clinical research.
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Radiotherapy modulates expression of EGFR, ERCC1 and p53 in cervical cancer

Cervical cancer is a public health problem and the molecular mechanisms underlying radioresistance are still poorly understood. Here, we evaluated the modulation of key molecules involved in cell proliferation, cell cycle and DNA repair in cervical cancer cell lines (CASKI and C33A) and in malignant tissues biopsied from 10 patients before and after radiotherapy. The expression patterns of epidermal growth factor receptor (EGFR), excision repair cross-complementation group 1 (ERCC1) and p53 were evaluated in cancer cell lines by quantitative PCR and western blotting, and in human malignant tissues by immunohistochemistry. The mutation status of TP53 gene was evaluated by direct sequencing. Among cell lines, absent or weak modulations of EGFR, ERCC1 and p53 were observed after exposure to 1.8 Gy. Conversely, increased expressions of p53 (5/10 patients; P=0.0239), ERCC1 (5/10 patients; P=0.0294) and EGFR (4/10 patients; P=0.1773) were observed in malignant tissues after radiotherapy with the same radiation dose. TP53 mutations were found only in one patient. Here we show that a single dose of radiotherapy induced EGFR, ERCC1 and p53 expression in malignant tissues from cervical cancer patients but not in cancer cell lines, highlighting the gap between in vitro and in vivo experimental models. Studies on larger patient cohorts are needed to allow an interpretation that an upregulation of p53, EGFR and ERCC1 may be part of a radioresistance mechanism.

Detection of EGFR Gene Mutation in Lung Adenocarcinoma by ARMS and Its Clinical Significance / 现代检验医学杂志

Objective To investigate the efficacy and clinical significance of amplification refractory mutation system (ARMS)in epidermal growth factor receptor (EGFR) gene mutation in lung adenocarcinoma.Methods Collected 566 specimens of lung adenocarcinomia in pathology from Department of the First Affiliated Hospital of Xi'an Jiaotong University from January 2015 to August 2016.As the research object,which included 34 cases of pleural cell specimens,401 cases of lung biopsy specimens and surgical specimens from 131 patients with ARMS to complete the above specimens EGFR gene mutation detection,analysis of EGFR gene mutations associated with non-small cell lung cancer patients clinical data.Results Among 566 cases of lung cancer specimens,the EGFR mutation rate of 239 cases of patients with smoking had no obvious correlation with age,gender and surgical methods(P＞0.05),but primary lung site was closely related (P＜0.05),and EGFR mutation rate of 327 cases of patients with non smoking had no obvious correlation withage,sex,operation mode and primary lung site (P＞0.05).Conclusion ARMS is an ideal method for the detection of EGFR gene mutation in non-small cell lung cancer.Smoking is a great influence on EGFR mutation rate in both lung tissue,and for right lung tissue is more dramatic.