肺癌细胞株中顺铂药物敏感性的相关基因

目的 筛选小细胞肺癌(SCLC)和非小细胞肺癌(NSCLC)细胞株中与顺铂(DDP)药物敏感性相关的基因.方法 采用二苯基溴化四氮唑蓝(MTT)比色分析法,测定4株SCLC细胞株和6株NSCLC细胞株对DDP的药物敏感性;应用cDNA微阵列技术检测肺癌细胞株中1291个药物敏感性相关基因的表达状态,并分析基因表达与DDP敏感性之间的相关性.结果 Metallothionein、Cathepsin B、TIMP1、TNF-R1、TGFβ-induced 68000、Cathepsin L、Galectin-1、Annexin 11、PAI-1、IGFBP4、UPAR、Jagged、CD13、α1 A-AR、EphA2(Eek)、APC、RhoC、Fibromodulin、GATA-6、HSC 70共20个基因,在SCLC和NSCLC细胞株中均与DDP的药物敏感性呈负相关,只有Procoagula和MDM2与DDP的敏感性呈正相关.VHL、MMP-7、Elongin A、GSK-3β、SLC、Galectin-3、Integrinβ5、Moesin、IKKβ和ETV 1等10个基因,只在SCLC细胞株中与DDP的药物敏感性呈负相关,而AT2与DDP敏感性呈正相关.Clusterin、FG FR-2、Thrombospondin 1、HSP 32、Lactate dehydrogenase A、P300、Thymosinβ10、CD81、C/EBPγ和Rak等10个基因,只在NSCLC细胞株中与DDP的药物敏感性呈负相关,而只有CaMKK和TPA与DDP的敏感性呈正相关.结论 与DDP药物敏感性相关的基因共有45个,其中共同表达于SCLC和NSCLC细胞株中的基因有22个,在SCLC细胞株中特异表达的基因有11个,在NSCLC细胞株中特异表达的基因有12个.     

人肺癌细胞中去甲长春新碱和泰索帝药物敏感性基因的比较分析

目的筛选人肺癌细胞株中与去甲长春新碱（NVB）、泰索帝（Doc）药物敏感性相关的基因。方法采用MTT比色分析法,测定4株小细胞肺癌（SCLC）细胞株和6株非小细胞肺癌（NSCLC）细胞株对NVB、Doc的药物敏感性;应用cDNA macroarray技术检测10株肺癌细胞株中1291个药物敏感性关联基因的表达状态,聚类分析二者间的相关性。结果（1）NVB对10株肺癌细胞株的作用效果明显优于Doc。（2）10株肺癌细胞株中,与Doc呈正相关的药物敏感性关联基因多于NVB,而与NVB呈负相关的多于Doc;而在6株NSCLC中,与Doc和NVB呈正相关、负相关一致的基因较多。（3）10株肺癌细胞株中,有51个基因与Doc、NVB药物敏感性相关联,占全部候选基因的3．95％（51／1291）。与NVB药物敏感性呈显著相关的基因27个,其中负相关24个,正相关3个;与Doc药物敏感性关联的基因24个,其中负相关13个,正相关11个。6株NSCLC株中,有67个基因与药物敏感性相关,占全部实验基因的5．19％（67／1291）。与NVB药物敏感性呈显著相关的基因29个,其中负相关25个,正相关4个;与Doc药物敏感性关联的基因38个,其中负相关34个．正相关4个。（4）Rab 1、Rab 3、Rho B、Rho C、Rac 1、Rac 2、Gho GDI B、CD44、Integrin α5、Integrin α6、Integrin β5、Vinculin等细胞骨架关联基因在SCLC、NSCLC中的表达存在差异。结论SCLC和NSCLC细胞株中,NVB、Doc药物敏感性相关联基因表达存在明显差异。     

肺癌细胞株中NVB和Doc药物敏感性关联基因的研究

目的:筛选小细胞肺癌(SCLC)和非小细胞肺癌(NSCLC)细胞株中与诺维苯(NVB)、泰索帝(Doc)药物感受性相关的基因.方法:采用MTT比色分析法测定4株SCLC细胞株和6株NSCLC细胞株对NVB、Doc的药物感受性,同时应用cDNA macroarray技术检测10株肺癌细胞株中1291个药物感受性关联基因的表达状态,分析二者之间的相关性.结果:与NVB和Doc药物敏感性关联(r≥±0.4)的基因共有56个.1)与NVB药物敏感性相关联的基因有36个,SCLC NSCLC组与NSCLC组共同表达的基因有20个,其中,SCLC NSCLC组表达的基因有27个,特异表达的有7个,NSCLC组为29个基因,9个基因特异表达.2)与Doc药物敏感性相关联的基因有50个,其中,SCLC NSCLC组与NSCLC组共同表达的基因有12个,SCLC NSCLC组表达的基因有24个,12个基因特异表达,NSCLC组为38个基因,26个基因特异表达.3)肺癌细胞株中与NVB、Doc药物感受性相关联基因的分类主要分布于:信号转导分子、代谢酶类及其抑制剂、细胞因子和转录因子等4类.结论:SCLC和NSCLC细胞株中NVB、Doc药物敏感性相关联基因存在明显差异,但分类基本相同.     

肺癌中EML4-ALK融合基因的存在状态及生物学功能

肺癌是当前世界上最常见的恶性肿瘤,主要包括小细胞肺癌(small cell lung cancer,SCLC)和非小细胞肺癌(non-small cell lung cancer,NSCLC)两类,而NSCLC约占肺癌总数的80%。尽管外科手术技术不断提高,化疗新药不断上市并进入临床应用,但肺癌患者的预后仍     

非小细胞肺癌细胞株抗癌药物敏感性相关基因差异表达的初步研究

背景与目的为了提高晚期肺癌的化疗效果，实行个体化治疗，筛选和鉴定肺癌细胞抗癌药物敏感性基因具有重要临床意义。本研究比较了非小细胞肺癌(NSCLC)和永生化人支气管上皮细胞(BET2A)细胞株抗癌药物敏感性相关基因的差异表达，以寻找与抗癌药物敏感性相关的基因。方法应用cDNA macroarray技术，对6株NSCLC和BET2A细胞株的抗癌药物敏感性相关基因进行差异表达分析，RT-PCR技术验证滤膜杂交结果。结果在1291个候选基因中筛选出73个差异表达基因，其中45个基因表达上调，28个基因表达下调。RT-PCR验证结果与cDNA macroarray检测结果一致。结论抗癌药物敏感性相关基因的差异表达可能是药物敏感性产生不同的原因之一。本研究结果为逆转肺癌多药耐药性提供了理论基础和新靶点，并为临床新药开发及个体化治疗提供了实验依据。     

2010年美国临床肿瘤学会年会热点——小细胞肺癌的研究进展

从生物学行为的角度可将肺癌分为小细胞肺癌(small cell lung cancer,SCLC)和非小细胞肺癌(non-small cell lung cancer,NSCLC),其中SCLC约占15%左右.SCLC恶性程度高,虽然对放疗、化疗较敏感,但预后差[1].本文就2010年美国临床肿瘤学会(American Society of Clinical Oncology,ASCO)年会上有关SCLC的研究进展做一综述.     

肺鳞状细胞癌的遗传学特点及靶向治疗进展

肺癌是全世界癌症死亡的首要原因,在我国,肺癌也是发病率和死亡率最高的恶性肿瘤。肺癌按组织病理学主要分成两大类:小细胞肺癌(small cell lung cancer,SCLC)和非小细胞肺癌(non-small cell lung cancer,NSCLC)。NSCLC主要又分成鳞状细胞癌(squamous cell carcinoma,SQCC)、腺状细胞癌(adenocarcinoma,A DC)和大细胞癌     

转化性小细胞肺癌1例分析

我国肺癌发病率及死亡率在恶性肿瘤中居首位[1].肺癌分为小细胞肺癌( small cell lung cancer,SCLC)和非小细胞肺癌(non -small cell lung cancer,NSCLC)[2].NSCLC 包括腺癌、鳞癌及大细胞肺癌.最常见的组织学分类是腺癌,其次是小细胞癌,鳞状细胞癌或神经内分...     

阿法替尼在NSCLC中的应用与展望

在全球肿瘤病死率中,肺癌病死率居首,5年生存率很低,只有15%。肺癌主要分为非小细胞肺癌（non-small-cell lung cancer,NSCLC）、小细胞肺癌（small cell lung cancer,SCLC）。NSCLC最常见,约占肺癌85%。     

吉西他滨联合顺铂治疗晚期非小细胞肺癌临床观察

我们采用吉西他滨(GEM)加顺铂(DDP)(GP方案)治疗ⅢB、Ⅳ期非小细胞肺癌(non-small cell lung cancer，NSCLC)患者20例，密切观察其临床近期疗效和毒副反应，现报告如下。     

In vivo screening models of cisplatin-resistant human lung cancer cell lines using SCID mice

In vivo screening models of a cisplatin (CDDP)-resistant human small-cell lung cancer cell (SCLC) line, H69/CDDP, and a non-small-cell lung cancer cell (NSCLC) line, PC-14/CDDP, were evaluated. The transplantability of the tumor xenografts to SCID mice was more than 90%. Tumor xenografts of H69/CDDP and PC-14/CDDP showed CDDP resistance during in vivo treatment. The novel anticancer agent 254-S showed only a partial effect on the growth of H69/CDDP and PC-14/CDDP while ormaplatin showed no cross resistance to CDDP. The in vivo results correlated well with the results of the in vitro MTT assay. In this in vivo sensitivity test, H69/CDDP and PC-14/CDDP were more sensitive to ormaplatin than its parental cell lines. In vivo sensitivity testing using SCID mice bearing transplanted CDDP-resistant tumors was shown to be useful for evaluating the effects of new anti-cancer drugs, especially those that might overcome CDDP resistance.Abbreviations SCLC small-cell lung cancer - NSCLC non-small-cell lung cancer - SCID severe combined immunodeficiency - CDDP cisplatin - PBS phosphate-buffered saline - FBS fetal bovine serum This work was supported, in part, by Grants-in-Aid for Cancer Research from the Ministry of Health and Welfare, the Comprehensive Ten-year Strategy for Cancer Control from the Ministry of Education, Science and Culture. Support from the Bristol-Myers Squibb Foundation is also appreciated     

An anti-GD2 monoclonal antibody enhances apoptotic effects of anti-cancer drugs against small cell lung cancer cells via JNK (c-Jun terminal kinase) activation.

Small cell lung cancer (SCLC) cell lines specifically express ganglioside GD2, and anti-GD2 monoclonal antibodies (mAbs) caused suppression of cell growth and induced apoptosis of SCLC cells with single use. Here, enhancement of the cytotoxic effects of various anti-cancer drugs with an anti-GD2 mAb was demonstrated. The cytotoxicity of all six drugs examined was markedly enhanced, i.e. 2.4 - 7.8-fold increase of cell sensitivity in terms of IC(50). In particular, the combination of cisplatin (CDDP) with an anti-GD2 mAb resulted in prominent enhancement of cytotoxicity even in low - moderate GD2-expressing lines. The anti-GD2 mAb induced weak activation of c-Jun terminal kinase (JNK) in SCLC cells, and all anti-cancer drugs also induced its activation to various degrees. When CDDP and an anti-GD2 mAb were used together, significantly stronger JNK activation was observed corresponding to the cytotoxic effects, suggesting that synergistic phosphorylation of JNK with two reagents induced prominent apoptosis. The essential role of JNK in the induction of SCLC apoptosis with CDDP and anti-GD2 mAb was confirmed by experiments with a JNK inhibitor, curcumin. These results suggest that anti-GD2 mAbs would be very efficient in combination with anti-cancer drugs, both to achieve SCLC-specific cytotoxicity and to enhance its magnitude.     

An Anti-GD2 Monoclonal Antibody Enhances Apoptotic Effects of Anti-cancer Drugs against Small Cell Lung Cancer Cells via JNK (c-Jun Terminal Kinase) Activation

Small cell lung cancer (SCLC) cell lines specifically express ganglioside GD2, and anti-GD2 monoclonal antibodies (mAbs) caused suppression of cell growth and induced apoptosis of SCLC cells with single use. Here, enhancement of the cytotoxic effects of various anti-cancer drugs with an anti-GD2 mAb was demonstrated. The cytotoxicity of all six drugs examined was markedly enhanced, i.e. 2.4–7.8–fold increase of cell sensitivity in terms of IC₅₀. In particular, the combination of cisplatin (CDDP) with an anti-GD2 mAb resulted in prominent enhancement of cytotoxicity even in low-moderate GD2–expressing lines. The anti-GD2 mAb induced weak activation of c-Jun terminal kinase (JNK) in SCLC cells, and all anti-cancer drugs also induced its activation to various degrees. When CDDP and an anti-GD2 mAb were used together, significantly stronger JNK activation was observed corresponding to the cytotoxic effects, suggesting that synergistic phosphorylation of JNK with two reagents induced prominent apoptosis. The essential role of JNK in the induction of SCLC apoptosis with CDDP and anti-GD2 mAb was confirmed by experiments with a JNK inhibitor, curcumin. These results suggest that anti-GD2 mAbs would be very efficient in combination with anti-cancer drugs, both to achieve SCLC-specific cytotoxicity and to enhance its magnitude.     

Chemosensitivity testing of human lung cancer cell lines using the MTT assay

Thirty human lung cancer cell lines were tested for chemosensitivity using the semi-automated, non-clonogenic MTT assay. The tumour cell lines came from three major categories of patients: untreated small cell lung cancer (SCLC); SCLC relapsing on chemotherapy; and non-SCLC predominantly from untreated patients. From these data IC50 values were derived for each drug in each cell line. While some inter-experimental variability was observed, the rank order of chemosensitivity of each cell line within this panel was significantly correlated between experiments. These results show that tumour cell lines derived from untreated small cell lung cancer patients were the most chemosensitive for adriamycin, melphalan, vincristine and VP16 compared to the other cell types. In addition, untreated SCLC was more sensitive than non-SCLC to BCNU and cis-platin, while vincristine was the only drug to which treated SCLC was more sensitive compared to the non-SCLC lines. In contrast, no significant differences between the lung cancer types were observed for vinblastine. Thus, this panel of lung cancer cells exhibited a drug sensitivity profile paralleling that observed in clinical practice. These results suggest that this lung cancer cell line panel in combination with a relatively simple but reproducible chemosensitivity assay, such as the MTT assay, has potential for the testing of drug combinations and evaluating new anti-cancer agents in vitro.     

Determinants of response to the DNA topoisomerase II inhibitors doxorubicin and etoposide in human lung cancer cell lines.

BACKGROUND: Small-cell lung cancer (SCLC) is more sensitive to anticancer agents than non-small-cell lung cancer (NSCLC), but few studies have analyzed the mechanisms of natural drug resistance responsible for this difference. PURPOSE: To elucidate these mechanisms, we determined drug sensitivity and evaluated the biochemical parameters affecting response to the DNA topoisomerase II inhibitors doxorubicin and etoposide in both types of cancer cell lines, in particular the activity and content of DNA topoisomerase II, as well as etoposide uptake and cell doubling time. METHODS: Drug sensitivity and cellular uptake of etoposide were determined by clonogenic assay and accumulation of radiolabeled drug, respectively. The topoisomerase II activity was assayed by decatenation of kinetoplast DNA to minicircle DNA using nuclear protein, and the content was determined by immunoblot analysis of nuclear extracts. We also compared the topoisomerase II content in parent cell lines with that in lines with cisplatin resistance acquired in vitro. RESULTS: Sensitivities to doxorubicin and etoposide were higher in SCLC cell lines than in NSCLC lines, and the difference was statistically significant. Etoposide uptake in SCLC cells was higher than in NSCLC cells; the difference was statistically significant, but this difference may not be sufficient to account for the variation in sensitivities of the cell lines. Topoisomerase II activities of nuclear protein from SCLC cell lines were reproducibly twofold higher than those for NSCLC cell lines. The topoisomerase II content in nuclear protein appeared to be higher in SCLC cell lines than in NSCLC cell lines and corresponded to the sensitivities to doxorubicin and etoposide. In the cisplatin-resistant NSCLC cell lines PC-7/CDDP and PC-14/CDDP, the topoisomerase II content was increased compared with that in the parent lines, but the topoisomerase II content in other cisplatin-sensitive parent lines was similar to that in resistant sublines. CONCLUSIONS: These findings suggest that the topoisomerase II activity and content may be major factors in determining sensitivity to topoisomerase II inhibitors.     

Levels of glutathione S transferase pi mRNA in human lung cancer cell lines correlate with the resistance to cisplatin and carboplatin

The amounts of mRNA for glutathione S transferase pi (GST pi) were significantly lower in 3 human small cell lung cancer (SCLC) cell lines than in 3 non small cell lung cancer (NSCLC) cell lines. The sensitivities of the 3 SCLC cell lines to cisplatin and carboplatin were much higher than those of the 3 NSCLC cell lines. These results indicate that low levels of GST pi mRNA expression in SCLC cell lines inversely correlate to high sensitivity to cisplatin and carboplatin, and further suggest that GST pi may play an important role in intracellular inactivation of these drugs.     

Chemosensitivity testing of small cell lung cancer using the MTT assay.

A simple colorimetric test, the MTT assay, has been adapted for chemosensitivity testing of human small cell lung cancer cell lines, and fresh tumour samples. Optimal conditions for clinical chemosensitivity testing were determined using established SCLC lines. Nineteen different chemotherapeutic agents were tested, and sixteen of them were found to be cytotoxic in this assay system. The drug sensitivity of a panel of 16 SCLC cell lines was measured and compared. There was very little intraexperiment variation, but the interexperiment variation was significant. Cell lines which were derived from patients who had not received chemotherapy at the time the cell line was established were more sensitive (to all but one of the drugs) than lines derived from treated patients, and the differences were statistically significant for two of the drugs. One cell line, NCI-H209, which was derived from an untreated patient, stood out as being the most sensitive or among the most sensitive to all of the drugs tested. Another cell line, H69AR, which is a multidrug resistant subline of the cell line NCI-H69, was the most resistant to many of the natural product drugs tested. Multiple drug chemosensitivity testing was performed on eight fresh tumour samples from SCLC patients (five pleural effusions, one lymph node, and two primary tumours). It was possible to perform chemosensitivity testing on all of the clinical samples in which sufficient tumour cells were available. The drug sensitivity of the clinical samples was, in most cases, within the same range as for the cell lines. Since this assay is very rapid and simple to perform, it may have practical applications in clinical drug sensitivity testing of human tumours.     

LEVELS OF GLUTATHIONES TRANSFERASE π mRNA IN HUMAN LUNG CANCER CELL LINES CORRELATE WITH THE RESISTANCE TO CISPLATIN AND CARBOPLATIN

The amounts of mRNA for glutathione S transferase π (GST π) were significantly lower in 3 human small cell lung cancer (SCLC) cell lines than in 3 non small cell lung cancer (NSCLC) cell lines. The sensitivities of the 3 SCLC cell lines to cisplatin and carboplatin were much higher than those of the 3 NSCLC cell lines. These results indicate that low levels of GST π mRNA expression in SCLC cell lines inversely correlate to high sensitivity to cisplatin and carboplatin, and further suggest that GST π may play an important role in intracellular inactivation of these drugs.     

Significance of nuclear glutathione S-transferase pi in resistance to anti-cancer drugs.

Recent study has shown that nuclear glutathione S-transferase (GST) pi accumulates in cancer cells resistant to doxorubicin hydrochloride (DOX) and may function to prevent nuclear DNA damage caused by DOX (Goto et al., FASEB J., 15, 2702 - 2714 (2001)). It is not clear if the amount of nuclear GSTpi increases in response to other anti-cancer drugs and if so, what is the physiological significance of the nuclear transfer of GSTpi in the acquisition of drug-resistance in cancer cells. In the present study, we employed three cancer cell lines, HCT8 human colonic cancer cells, A549 human lung adenocarcinoma cells, and T98G human glioblastoma cells. We estimated the nuclear transfer of GSTpi induced by the anti-cancer drugs cisplatin (CDDP), irinotecan hydrochloride (CPT-11), etoposide (VP-16) and 5-fluorouracil (5-FU). It was found that: (1) Nuclear GSTpi accumulated in these cancer cells in response to CDDP, DOX, CPT-11, VP-16 and 5-FU. (2) An inhibitor of the nuclear transport of GSTpi, edible mushroom lectin (Agaricus bisporus lectin, ABL), increased the sensitivity of the cancer cells to DOX and CDDP, and partially to CPT-11. Treatment with ABL had no apparent effect on the cytotoxicity of VP-16 and 5-FU. These results suggest that inhibitors of the nuclear transfer of GSTpi have practical value in producing an increase of sensitivity to DOX, CDDP and CPT-11.     

Additive and differential biological activity of alpha-interferon A, difluoromethylornithine, and their combination on established human lung cancer cell lines

The effect of human recombinant leukocyte interferon A (IFN-alpha A) and DL-alpha-difluoromethylornithine (DFMO) as single drugs and in combination on the in vitro growth, cell cycle distribution, activity of the enzyme L-dopa decarboxylase, and expression of the c-myc and N-myc oncogenes was studied in human lung cancer cell lines. In vitro growth activities were tested in concentrations ranging from 10 to 50,000 IU/ml for IFN-alpha A and from 0.1 to 10 mM for DFMO by means of the soft agarose clonogenic assay using continuous drug exposure. Ten well established small cell lung cancer (SCLC) cell lines including five cell lines of the classic and five of the variant phenotype, two cell lines derived from adenocarcinoma of the lung, and one large cell lung cancer cell line were included in the study. We found that IFN-alpha A inhibited the growth only of the variant phenotype of SCLC with an approximate drug concentration yielding a 50% inhibition of colony growth of 1000 IU/ml. None of the SCLC classic cell lines was inhibited significantly. The growth inhibition of IFN-alpha A correlated with the proliferation rate of the tumor. IFN-alpha A inhibited one of two adenocarcinoma cell lines and 0 of 1 large cell lung cancer cell line. DFMO inhibited the colony formation of 10 of 10 SCLC cell lines, 2 of 2 adenocarcinoma cell lines, and 0 of 1 large cell lung cancer cell line with a drug concentration yielding a 50% inhibition of colony growth of 1 mM. No difference between the classic and variant phenotypes of SCLC was found. The combination of IFN-alpha A and DFMO resulted in an additive cytostatic effect in all cell lines tested. The same result, i.e., an additive cytostatic effect, was obtained for two SCLC cell lines that were tested in liquid culture. Neither single drugs nor their combination led to an accumulation of cells in a particular phase of the cell cycle nor did it affect the activity of the SCLC classic marker enzyme L-dopa decarboxylase. In addition, IFN-alpha A, DFMO, and their combination did not affect the expression of the c-myc and N-myc oncogenes in cell lines NCI-N417 and NCI-H526, respectively, following 4, 24, and 72 h of continuous drug exposure.(ABSTRACT TRUNCATED AT 400 WORDS)