Clinical Course Of Patients with Small Cell Lung Cancer As Second Primary Malignancy

Objective： To evaluate the clinical course of patients with small cell lung cancer （SCLC） as second primary malignancy. Methods： Among the 355 patients diagnosed with SCLC at Helen and Harry Gray Cancer Center of Hartford Hospital Connecticut USA between 1988 and 1998, the records of 48 patients, which had been diagnosed with other malignancies before their diagnosis of SCLC, were retro- spectively reviewed. Results： Forty-eight patients （13.5%） were diagnosed with other malignancies prior to their SCLC among which 43 had documented smoking history and 93% of them （40/43） were current/former smokers. Of the 28-second primary SCLC patients who were treated with standard method, 11 （39.3%） achieved CR. 12 （42.8%） achieved PR, and the RR was 82.1%. The median survival of the 28 treated with standard method was 11.3 months （5.1-77.7 months）, while that of the rest 19 untreated patients （1 of 20 was lost to follow-up） was only 2.0 months （0.5 34.0 months）. There was no significant difference in the median survival and RR between 165 treated first primary SCLC （13.5 months and 77.6% respectively） and 28 treated secondary primary SCLC （11.3 months and 82.1% respectively） （P〉0.05）. The patients who had prostate cancer were older and subjected to less treatments than those with skin cancer, so their survival was shorter than the latter （3.5 months vs. 15 months, P〈0.05）. Conclusion： The response and survival of the treated patients with SCLC as a second malignancy showed no difference as compared to the treated ones with SCLC only. Therefore, an active medical treatment is important to relieve symptom and prolong survival of the second primary SCLC patients.

DIAGNOSTIC VALUE OF SERUM PROGRP31-98 IN PATIENTS WITH SMALL-CELL LUNG CANCER

Objective To explore the clinical significance of serum level of pro gastrin releasing peptide 31 98 (ProGRP31 98) for small cell lung cancer (SCLC), in comparison with neuron specific enolase (NSE). Methods Serum level of ProGRP31 98 and NSE was measured by ELISA respectively in 30 patients with SCLC, 30 with non small cell lung cancer (NSCLC), 15 with benign lung diseases and 15 normal subjects, additionally, 10 SCLC patients after having treatment with chemotherapy were included. The receiver operating characteristic (ROC) curve was used to set the cut off value and evaluate the diagnostic accuracy. Results The serum level of ProGRP31 98 was higher in patients with SCLC than that in other groups. The SCLC patients with extensive disease had a higher value than the patients with limited disease. In SCLC patients with distant metastases, it was also higher than in those without. Increase in serum ProGRP31 98 and NSE was both seen in SCLC patients, but for the former one, the increase was of much greater compared to the normal controls. Given the cut off value for ProGRP31 98 was 40ng·L -1 and for NSE 8μg·L -1 , their sensitivity of diagnosis in SCLC was 73% and 60%, respectively. The area under ROC curve of ProGRP31 98 was significantly larger than that of NSE. All patients responded to chemotherapy showed marked decrease in ProGRP31 98. Conclusion ProGRP31 98 is a more specific and sensitive marker than NSE in the diagnosis of SCLC.

HYPERMETHYLATION OF p14^(ARF) PROMOTER REGION AND EXPRESION OF p14^(ARF) GENE PRODUCT IN NON-SMALL CELL LUNG CANCER

Objective： This study was designed to investigate promoter methylation status and protein expression of p14^ARF gene in non-small cell lung cancer, and value the role of p14^ARF promoter methylation in carcinogenesis of non-small cell lung cancer. Methods： Promoter methylation status and protein expression of p14^ARF gene in 40 cases of non-small cell lung cancer were analyzed by methylation specific polymerase china reaction （MSP）, restriction enzyme-related polymerase chain reaction （RE-PCR） and immunohistochemistry （IHC）. Results： The positive rates of p14^ARF promoter methylation in tumor tissues and normal tissues adjacent to cancer were 17.5% （7/40） and 2.5% （1/40） respectively. There were statistically significant differences between them, P〈0.05. The results of RE-PCR were consistent with that of MSP. The expression rate of p14^ARF protein in tumor tissues was significantly lower than that in normal tissues adjacent to cancer, p〈0.01. Promoter methylation status and protein expression of p14^ARF gene in non-small cell lung cancer showed significantly an inverse correlation （r=-0.56, P〈0.01）, and both of them did not relate statistically with the clinicopathologic characteristics of patients such as histological classification, clinical stage, differentiation grade and lymph node involvement. Conclusion： Promoter methylation is a crucial mechanism of inactivation of p14^ARF gene. Promoter methylation of p14^ARF gene might he involved in carcinogenesis of non-small cell lung cancer, and is an early event in development process of non-small cell lung cancer. It might be used as a new target in gene treatments in the future.

Therapy-related myeloid leukemia during erlotinib treatment in a non-small cell lung cancer patient:A case report

BACKGROUND Epidermal growth factor receptor tyrosine kinase inhibitors(EGFR-TKIs)are tolerable drugs used for patients with EGFR-mutant advanced non-small cell lung cancer(NSCLC).Serious adverse reactions are uncommon compared with cytotoxic drugs.CASE SUMMARY A 52-year-old man presented with general weakness and cytopenia.He had been taking erlotinib for 11 mo to treat NSCLC.The pathological diagnosis from the right upper lobe mass was adenocarcinoma with an EGFR mutation in exon 21(L858R).He had previously received paclitaxel/carboplatin,gemcitabin/vinorelbine chemotherapy,stereotactic radiosurgery for brain metastasis,and whole-brain radiotherapy as treatment for NSCLC.We diagnosed the patient with acute myeloid leukemia(AML).During the induction and consolidation chemotherapy for AML,the erlotinib was discontinued.When complete remission of the AML was achieved,since the lung masses were increased,pemetrexed/cisplatin for the NSCLC was initiated.After two cycles of chemotherapy,the cytopenia was prolonged.AML relapse occurred with the same karyotype.CONCLUSION Therapy-related acute myeloid neoplasm(t-MN)is a rare but fatal late complication.Although a patient may be taking EGFR-TKIs,the possibility of t-MN should be considered.Further studies are needed to determine whether EGFR-TKI usage is a predisposing factor for t-MN.

EXPRESSION OF FRAGILE HISTIDINE TRIAD AND P53 IN NON-SMALL CELL LUNG CANCER

Objective： To investigate the expression offragile histidine triad （FHIT） and p53 protein in non-small cell lung cancer （NSCLC） and explore the relationship between their expressions and the clinicopathological features. Methods： FHIT protein and p53 protein were detected by immunohistochemistry in 76 cases of NSCLCs and matched normal lung tissues. Results： Fifty-one cases （67.1%） showed negative expression of FHIT （apparent reduction or loss） and thirty-seven cases （48.7%） showed p53 positive expression （overexpression）. The difference was significant （P=0.04）. However, there was no significant difference in FHIT expression between the p53-positive group and the p53-negative group （64.9% versus 69.2%, P=0.686）. The negative rate of FHIT protein expression was higher in squamous cell carcinoma than in adenocarcinoma, in moderately and poorly differentiated carcinoma than in well-differentiated carcinoma, and in cases with smoking history than in cases without smoking history （P〈0.05）. There was no relationship between FHIT expression and clinical stage or lymph node metastasis. The negative FHIT expression was not an independent predictor of overall survival （P=0.338）. Conclusion： The frequency of negative expression of FHIT protein is higher than that of positive expression of p53 in NSCLCs. The negative expression of FHIT is independent of the expression of p53. The change of expression of FHIT may play a role in the smoking related lung tumorigenesis while it may have no relationship with the progress of NSCLC or prognosis of the patients.

Combination chemotherapy with irinotecan and cisplatin as second-line treatment for small cell lung cancer

Objective： To evaluate the efficacy and safety of irinotecan （CPT-11） plus cisplatin （DDP） in patients with small cell lung cancer （SCLC） as second-line chemotherapy. Methods： Patients received irinotecan 60 mg/m^2 on days 1, 8, 15, and cisplatin 25 mg/m^2 on days 1-3, every 28 days one cycle. Response was evaluated every two cycles and patients were follow-up for two years or until death. Results： Among the 28 evaluable patients, there were 1 CR, 7 PR, 8 SD and 12 PD. The response rate was 28.6% （8/28）. Median time to progression （TTP） was 3.2 （0.8-5.6） months. Median survival after second-line treatment was 7.5 （1.5-31） months and overall survival was 15 （2.3-43.5） months. The most common adverse effect was hematological toxicity with 36.7% （11/30）, grades Ⅲ-Ⅳ neutroperia. Hepatic toxicity was another major side effect. Only one patient developed grade Ⅲ diarrhea. Conclusion： The combination of irinotecan and cisplatin is feasible, effective, and safe for SCLC as second-line treatment.

Role of positron emission tomography-computed tomography in non-small cell lung cancer

Lung cancer is the leading cause of cancer-related mortality worldwide. Non-small cell carcinoma and small cell carcinoma are the main histological subtypes and constitutes around 85% and 15% of all lung cancer respectively. Multimodality treatment plays a key role in the successful management of lung cancer depending upon the histological subtype, stage of disease, and performance status. Imaging modalities play an important role in the diagnosis and accurate staging of the disease, in assessing the response to neoadjuvant therapy, and in the follow-up of the patients. Last decade has witnessed voluminous upsurge in the use of positron emission tomography-computed tomography(PET-CT); role of PET-CT has widened exponentially in the management of lung cancer. The present article reviews the role of 18-fluoro-deoxyglucose PET-CT in the management of non small cell lung cancer with emphasis on staging of the disease and the assessment of response to neoadjuvant therapy based on available literature.

Gastric metastasis of small cell lung carcinoma:Three case reports and review of literature

BACKGROUND Small cell lung carcinoma(SCLC)is highly susceptible to metastasis in the early stages of the disease.However,the stomach is an uncommon site of metastasis in SCLC,and only a few cases of this type of metastasis have been reported.Therefore,SCLC gastric metastases have not been systematically characterized and are easily missed and misdiagnosed.CASE SUMMARY We report three cases of gastric metastasis from SCLC in this article.The first patient presented primarily with cough,hemoptysis,and epigastric fullness.The other two patients presented primarily with abdominal discomfort,epigastric distension,and pain.All patients underwent gastroscopy and imaging examinations.Meanwhile,the immunohistochemical results of the lesions in three patients were suggestive of small cell carcinoma.Finally,the three patients were diagnosed with gastric metastasis of SCLC through a comprehensive analysis.The three patients did not receive appropriate treatment and died within a short time.CONCLUSION Here,we focused on summarizing the characteristics of gastric metastasis of SCLC to enhance clinicians'understanding of this disease.

ANTI-HUMAN LUNG GIANT CELL CANCER (PG) EFFECT OF HUMAN LAK CELLS IN VITRO AND IN NUDE MICE

Human LAK cells were prepared by culturing normal human peripheral blood mononuclear cells (PBMC) with or without rIL-2 and assayed for T cell surface markers as well as anti-tumor activity against PC in vitro and in nude mice. Although the percentages of T3, T4, and T8 positive cells in rIL-2-activated cells did not differ significantly from those of control cells in vitro, the former showed stronger cytotoxicity than control cells to PG tumor cells in vitro. In vivo, LAK cells completely inhibited the growth of PG tumor in nude mice, whereas PBMC control cells were to be of no effect. The anti-tumor effect of human LAK cells in nude mice may offer a useful model to study the role of human LAK cells against human tumor in vivo.

PRDM14 Promotes the Migration of Human Non-small Cell Lung Cancer Through Extracellular Matrix Degradation in vitro

Background： As a novel molecular markerof non-small cell lung cancer （NSCLC）, PRDI-BFI and R1Z homology domain containing protein 14 （PRDM 14） is over-expressed ill NSCLC tumor tissues. Extracellular matrix degradation mediated by the balance between matrix metalloproteinases （MMPs） and tissue inhibitor of metalloproteinases （TIMPs） is one of the most important mechanism in king cancer metastasis. This study aimed to determine if PRDM14 promoted the migration of NSCLC cells through extracellular matrix degradation mediated by change of MMP/TIMP expression. Methods： The expression of PRDM 14 was down-regulated in human cell line A 549 after transfection with lentiviral vector-mediated short-hairpin ribonucleic acids （shRNAs） which targeted the PRDM 14 promoter. Cellular migration ofshRNA-infected cells was detected by a scratch wound healing assay and transwell cell rnigration assay. Expression levels of MMP1, MMP2, TIMP1, and TIMP2 were measured by quantitative real-time polymerase chain reaction （RT-PCR）. Results： Migration of PRDM 14-shRNA-infected cells was significantly inhibited relative to control cells as measured by the scratch wound healing （P 〈 0.05） and transwell cell migration assays （P 〈 0.01 ）. The expression of MMPI in A549 cells infected by PRDMI4-shRNA was down-regulated significantly （P 〈 0.01 ）, whereas the expression ofTIMP 1 and TIMP2 was up-regulated significantly （P 〈 0.01 ）. Conclusions： PRDM 14 accelerates A549 cells migration in vitro through extracellular matrix degradation. PRDM 14 is considered as a potential therapeutic target in metastatic NSCLC.

Over-expression of small ubiquitin-like modifier proteases 1 predicts chemo-sensitivity and poor survival in non-small cell lung cancer

Background Non-small cell lung cancer （NSCLC） is one of the most common malignant tumors.Despite the advances in therapy over the years,its mortality remains high.The aim of this study was to evaluate the expression of small ubiquitin-like modifier （SUMO） proteases 1 （SENP1） in NSCLC tissues and its role in the regulation of vascular endothelial growth factor （VEGF） expression.We also investigated the association between the expression level of SENP1 and the clinicopathological features and survival of the patients.Methods A SENP1 small interfering RNA （siRNA） was constructed and transfected into the NSCLC cells.VEGF gene expression was analyzed by real-time polymerase chain reaction （RT-PCR）.Immunohistochemistry staining was used to assess the expression of SENP1 in 100 NSCLC patients and its association with the clinicopathological features and survival was analyzed.Results VEGF expression was significantly higher in NSCLC tissues than in normal lung tissues.Inhibition of SENP1 by siRNA was associated with decreased VEGF expression.SENP1 was over-expressed in 55 of the 100 NSCLC samples （55％） and was associated with a moderate and low histological tumor grade （3.6％,38.2％,and 58.2％ in high,moderate and low differentiated tumors,respectively,P=0.046）,higher T stage （10.9％ in T1,and 89.1％ in T2 and T3 tumor samples,P ＜0.001）and TNM stage （10.9％ in stage Ⅰ,and 89.1％ in stages Ⅱ and Ⅲ tumor samples,P ＜0.001）.The rate of lymph node metastasis was significantly higher in the SENP1 over-expression group （76.4％） than that in the SENP1 low expression group （33.3％,P ＜0.001）.Sixty three patients received postoperative chemotherapy,including 34 with SENP1 over-expression and 29 with SENP1 low expression.Among the 34 patients with SENP1 over-expression,22 （64.7％） patients developed recurrence or metastasis,significantly higher than those in the low expression group 27.6％ （8/29） （P=0.005）.Multivariate Cox regression analysis showed that lymph node metastasis （P=0.015）,TNM stage （P=-0.001）,and SENP1 expression level （P=0.002） were independent prognostic factors for the survival of NSCLC patients.Conclusions SENP1 may be a promising predictor of survival,a predictive factor of chemo-sensitivity for NSCLC patients,and potentially a desirable drug target for lung carcinoma target therapy.

Acute myocardial infarction mimicking squamous cell lung cancer with bone metastases due to hypercalcemia： a case report

Acute myocardial infarction （AMI）, the most severe .coronary artery disease, is one of the most frequent cardiac emergencies, and early diagnosis and treatment are very important to decrease the subsequent cardiac adverse events such as malignant arrhythmia and sudden cardiac death. But in fact, lots of diseases are similar to AMI in clinical practice, of which the most common are myocarditis, pulmonary embolism in department of cardiology. Here we report a case of AMI-like squamous cell lung cancer with bone metastases.

Lung carcinoma with spindle and/or giant cell： a clinicopathological analysis of 17 cases

Objective： Lung carcinoma with spindle and （or） giant cell （LCSG） is a rare epithelial malignant tumor. The aim of our study is to investigate the clinicopathological and prognostic characteristics of 17 cases of LCSGs. Methods： Among 421 patients underwent resection of lung carcinomas, 17 cases of LCSG were studied for clinical, gross and histological parameters. Follow-up information was obtained and analyzed to clarify prognostically significant parameters. Results： The LCSG patients consisted of 15 males and 2 females, with the age ranging from 45 to 78 years （median, 58 years）; 5 cases of stage Ⅰ, 3 of stage Ⅱ, 9 of stage Ⅲ by pathological TNM staging; 2 cases of exclusively spindle cell carcinoma, 5 cases of lung carcinoma with spindle cell, 10 cases of lung carcinoma with giant-cell carcinoma. Cough, chest distress, or chest pain were the most common presenting symptoms, occurring in 15 patients （88.2%）. Of 5 patients in stage Ⅰ, 4 were alive and free of relapse for more than 5 years. The difference in survival was statistically significant between LCSG and squamous cell carcinoma patients （median survival, 36 vs. 61 months; P = 0.027）. Lymph node metastasis and carcinoma with giant cell were the hazardous factors impacting postoperative prognosis of LCSG patients. Conclusion： LCSG patients in early stage may have an optimistic outcome. Lung carcinomas with giant cell displayed multiple cell components in histopathology, and poor outcome due to more lymph node involved.

可切除非小细胞肺癌新辅助免疫治疗研究进展

近年来,免疫检查点抑制剂(immune checkpoint inhibitors,ICIs)对晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)患者预后的改善已成为共识,越来越多的临床研究也逐渐证明了免疫治疗对于可切除NSCLC患者的重要价值。然而,目前关于新辅助治疗背景下免疫联合策略的探索、治疗相关副作用、预后生物标志物等问题仍存在争议。本文综述了可切除NSCLC患者新辅助免疫治疗的最新进展,引发了新的思考,并讨论了其在临床应用中的优势及挑战。

小细胞肺癌分子分型:向临床实践转化的挑战

小细胞肺癌(small cell lung cancer,SCLC)是肺癌的组织学亚型之一,特点是高增殖、早转移、易耐药和复发。数年来SCLC一直被视为一种同质性疾病,治疗采用统一的放化疗策略。尽管早期疗效显著,但耐药和复发很快出现,缺乏满意的治疗效果,这可能是因为目前对SCLC的肿瘤异质性认识不足所导致。最近,临床前研究提出了基于谱系转录因子相对高表达定义的SCLC分子分型概念。本文主要阐述SCLC分子分型的现状和相关最新发现,强调分子分型转化在临床实践中所遇到的问题,旨在增进对SCLC分子分型研究进展的认识。

系统免疫炎症指数对根治性放疗Ⅲ期肺鳞癌患者长期生存的预测价值

目的探讨系统免疫炎症指数(SII)对接受根治性放疗的Ⅲ期肺鳞癌患者长期生存的预测价值。方法回顾性收集接受根治性放疗的Ⅲ期肺鳞癌患者的临床资料。计算患者放疗前1周内的SII及相关炎症指标,同时应用X-Tile软件确定最佳截断值。分析患者的生存情况以及SII对患者总生存(OS)及无进展生存(PFS)的影响。结果共纳入了453例患者,低SII组336例(<1277.3),高SII组117例(≥1277.3)。高SII组的中位OS和中位PFS均较低SII组缩短(OS:20.8个月vs.31.0个月,Log-rankχ2=18.015,P<0.01;PFS:13.0个月vs.21.0个月,Log-rankχ2=15.062,P<0.01)。多因素Cox回归分析显示,高SII是患者OS(HR=1.628,95%CI:1.294~2.047,P<0.001)和PFS(HR=1.559,95%CI:1.240~1.961,P<0.001)的独立危险因素,其他的影响因素包括较晚的TNM分期、放疗疗效欠佳,HALP评分下降。结论SII可作为接受根治性放疗Ⅲ期肺鳞癌患者长期生存的评价指标,SII升高提示预后较差。

多倍体肿瘤巨细胞对肿瘤耐药作用机制的研究进展

化疗是肿瘤治疗的常用手段,在肿瘤治疗前期具有积极作用,但随着化疗疗程的增加,肿瘤细胞对化疗药物的敏感性逐步减弱,从而产生化疗耐药性。肿瘤耐药是多因素介导的复杂过程,目前认为其与肿瘤细胞干性、异质性、肿瘤免疫微环境和化疗药物转运与外排增加等有关,但具体的机制尚不清楚。多倍体肿瘤巨细胞(polyploid giant cancer cell,PGCC)是一类体积增大、胞核丰富的特殊肿瘤细胞亚群。研究发现PGCC普遍存在于结直肠癌、乳腺癌、卵巢癌等多种肿瘤中,与肿瘤的发生、转移、耐药和复发有着密切联系。阐述PGCC在肿瘤中的形成及意义,并从PGCC所具有的特殊机制、自噬、衰老和DNA修复等角度阐述PGCC引起耐药发生的可能机制,从而为改善肿瘤耐药提供可能策略。

汉防己乙素衍生物LYY-47对三阴性乳腺癌细胞及其多倍体巨瘤细胞增殖、凋亡的作用和机制

目的探讨汉防己乙素衍生物LYY-47对三阴性乳腺癌(TNBC)细胞及其多倍体巨瘤细胞(PGCCs)增殖、凋亡的作用和机制。方法取TNBC MDA-MB-231细胞和MDA-MB-436细胞培养至对数生长期,诱导形成PGCCs,培养35 d,采用苏木素-伊红(H&E)染色观察不同培养时间时2种细胞的PGCCs形态学特征并进行计数;收集MDA-MB-231细胞、PGCCs及其子代细胞,采用流式细胞仪检测分析细胞周期,采用Western blot检测周期相关蛋白[细胞周期蛋白依赖性激酶1(CDK1)和细胞周期蛋白B1(CyclinB1)]、干性相关蛋白[乙醛脱氢酶1A1(ALDH1A1)、白细胞分化抗原44(CD44)及白细胞分化抗原133(CD133)]、DNA损伤修复相关蛋白[布卢姆(BLM)、Rad51及乳腺癌易感基因1(BRCA1)]及凋亡相关蛋白[BCL2-相关X蛋白(Bax)、B淋巴细胞瘤蛋白-2(Bcl-2)、裂解凋亡蛋白酶-3(cleaved Caspase-3)及裂解凋亡蛋白酶-8(cleaved Caspase-8)]的表达,采用噻唑蓝(MTT)法和克隆形成实验检测细胞活力和细胞集落数,采用细胞免疫荧光实验检测磷酸化组蛋白2AX(γ-H2AX)的表达;采用荧光偏振实验检测BLM DNA解旋酶的活性;收集对数生长期MDA-MB-231细胞,分为对照组(同等体积的完全培养基)、紫杉醇(PTX)组(500 nmol/L PTX)、3-CF 3,4-F-苯基类似物(ML216)组(3μmol/L ML216)及ML216+PTX组(500 nmol/L PTX和3μmol/L ML216),采用Image J软件计数各组细胞数;收集对数生长期MDA-MB-231细胞及其PGCCs子代细胞,分为对照组(0.00μmol/L)、LYY-47给药组(2.50μmol/L、5.00μmol/L及6.50μmol/L),采用流式细胞仪检测上述各组细胞的凋亡情况;6周龄雌性无特定病原体(SPF)级无胸腺BALB/c裸鼠12只,皮下分别注射5×10^(6)个MDA-MB-231细胞及其PGCCs子代细胞,每隔3天测量1次肿瘤体积,连续29 d,处死裸鼠剥离肿瘤、称重,取肿瘤组织制作切片,采用H&E染色和免疫组织化学染色观察细胞形态和检测BLM、Ki-67的表达。结果与TNBC MDA-MB-231和MDA-MB-436细胞相比,PTX诱导的PGCCs出现增大的细胞核和细胞质区域,通过不对称分裂产生子代细胞;与MDA-MB-231细胞相比,PGCCs中S期和G2/M期细胞增加、CDK1和CyclinB1蛋白表达下调(P<0.05或P<0.01),其子代细胞中S期细胞增加、G2/M期细胞减少且CDK1、CyclinB1蛋白表达上调(P<0.05或P<0.01),PGCCs及其子代细胞中ALDH1A1、CD44及CD133蛋白表达上调(P<0.05),PGCCs子代细胞的增殖和克隆形成能力增强(P<0.05),γ-H2AX、BLM、BRCA1及Rad51蛋白表达上调(P<0.05);与PTX组相比,ML216+PTX组PGCCs形成的数量明显减少(P<0.01);PGCCs子代细胞体内成瘤的生长速度、肿瘤的体积和重量均大于MDA-MB-231细胞(P<0.01),瘤体中BLM与Ki-67均呈高表达(P<0.01);与对照组相比,LYY-47给药组BLM 642-1290 DNA解旋酶的ATPase、dsDNA解链活性以及DNA结合活性均下调(P<0.05),MDA-MB-231细胞及其PGCCs子代细胞中BLM、Rad51及BRCA1蛋白表达也均下调(P<0.05);与对照组相比,LYY-47给药组和ML216给药组MDA-MB-231及其PGCCs子代细胞增殖和克隆形成能力降低(P<0.05),LYY-47给药组能够引起MDA-MB-231及其PGCCs子代细胞G2/M期细胞增加(P<0.05)、S期细胞减少(P<0.05)、细胞内周期相关蛋白CDK1与CyclinB1的表达下调(P<0.05),ML216给药组MDA-MB-231及其PGCCs子代细胞G1期细胞增多、S期细胞减少(P<0.05);与对照组比较,LYY-47给药组MDA-MB-231细胞及其PGCCs子代细胞的总凋亡比例增加、Bcl-2表达下调及Bax、cleaved Caspase-3、cleaved Caspase-8蛋白表达上调(P<0.05)。结论LYY-47和ML216可影响TNBC及其PGCCs子代细胞的增殖,其机制可能与抑制BLM DNA解旋酶诱导细胞凋亡和阻滞细胞周期有关。

ITGA6基因对非小细胞肺癌脑转移瘤细胞的影响

目的深入探究ITGA6基因在非小细胞肺癌(non-small cell lung cancer,NSCLC)脑转移瘤细胞增殖、细胞周期调控、侵袭及迁移过程中的具体作用机制。方法通过挖掘GEO数据库中的NSCLC脑转移瘤相关数据,本研究成功识别出ITGA6基因为与NSCLC脑转移瘤发生和发展相关的重要因子。在此基础上,进一步探究该基因表达水平与NSCLC脑转移瘤患者预后之间的关联性。为验证ITGA6在NSCLC脑转移瘤中的表达模式,本研究进一步采用实时定量逆转录PCR(qRT-PCR)和Western Blot实验技术,在NSCLC脑转移瘤细胞系测定ITGA6表达水平。利用基因转染技术,在体外NSCLC脑转移瘤细胞系H1915-BrM和A549-BrM中分别转染阴性对照(NC组)和ITGA6特异性沉默载体(si-ITGA6组)。通过CCK-8增殖实验评估细胞增殖能力;利用EDU实验进一步验证细胞增殖变化;应用流式细胞技术检测细胞周期分布情况;通过Transwell侵袭实验和划痕愈合实验分析细胞侵袭和迁移能力。利用Western Blot技术检测上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白的表达水平。结果利用GEO数据库系统分析NSCLC脑转移瘤的差异表达基因,并发现ITGA6是其中一个关键的上调基因。临床数据分析表明:ITGA6高表达与NSCLC脑转移瘤患者不良预后显著相关(P<0.05)。进一步通过qRT-PCR和Western Blot技术在实验层面验证ITGA6在肺癌脑转移瘤细胞系中高表达。CCK-8和EDU实验结果表明:si-ITGA6显著抑制NSCLC脑转移瘤细胞H1915-BrM和A549-BrM的增殖(P<0.05)。流式细胞技术揭示:si-ITGA6处理后,细胞在G0/G1期比例增加,并且S期及G2期细胞随之减少,表明细胞周期被阻滞(P<0.05)。Transwell和划痕实验均显示,si-ITGA6显著降低NSCLC脑转移瘤细胞侵袭和迁移能力。Western Blot观察分析:si-ITGA6组中上皮标记物E钙黏蛋白表达显著上升,而间质标记物纤连蛋白、波形蛋白以及基质金属蛋白酶(MMP-2和MMP-9)表达则显著下降,提示EMT过程受到抑制。结论通过特异性沉默ITGA6基因,能够显著抑制NSCLC脑转移瘤细胞增殖、迁移和侵袭能力,同时阻滞细胞周期,并调控EMT相关蛋白的表达。

小细胞肺癌骨转移的机制及其诊断标志物的研究进展

小细胞肺癌(small cell lung cancer,SCLC)是一种恶性程度高、转变迅速、侵袭转移快的肺癌亚型,极易出现早期转移,预后较差。SCLC骨转移共经历癌细胞在原发部位增殖并突破局部组织、进入血液循环形成循环肿瘤细胞(circulating tumor cells,CTCs)并通过血液循环到达骨组织、到达骨组织后在骨微环境的支持下生根发芽形成新的肿瘤灶等三大阶段,而传统的影像学、病理学检查具有敏感性低、价格昂贵、实施难度大等缺点,基于血液标志物检测作为SCLC骨转移筛查与疗效评估的探索性研究近年多有报道。本文通过综述SCLC骨转移形成的分子生物学机制,发现SCLC骨转移中影像学及病理活检等常规诊断方法的不足,透明质酸、蛋白质生物标志物、非编码RNA、液体活检中的生物标志物等变化早于影像学变化,具有操作简便、可重复性好等优点,为SCLC骨转移早期诊断提供新思路、新方法,值得在临床推广应用。