The inhibitory effect of cisplatin in combination with irradiation on lung tumor cell growth is due to induction of tumor cell apoptosis

Chemoradiation is becoming an important method of treatment of advanced lung cancer. Although many studies have demonstrated the efficacy of chemoradiation against lung tumors, the mechanism of its effect is still poorly understood. In this study, we analyzed the combined effect of cisplatin (CDDP) and irradiation on human lung cancer cell lines (PC-9 and Lu 134A) in vitro using MTT assay and isobologram analysis. To study whether the combined effect is due to induction of tumor cell apoptosis, we further quantified the nuclear fragmentation of tumor cells by flow cytometric analysis. Our study revealed that a combination of cisplatin and irradiation had an additive inhibitory effect on both PC-9 (adenocarcinoma) and Lu 134A (small cell carcinoma) tumor cell growth. Cisplatin improved the sensitivity of tumor cells to irradiation. The use of both cisplatin and irradiation doses could be reduced without decreasing the inhibitory effect. The additive inhibitory effect was correlated positively with the levels of tumor cell nuclear fragmentation. We concluded that combination treatment with cisplatin and irradiation is effective against lung tumors, not only small cell carcinoma but also non-small cell carcinoma, and the inhibitory effect was due to induction of tumor cell apoptosis.     

Efficacy of 6-diazo-5-oxo-L-norleucine and N-[N-gamma-glutamyl-6-diazo-5-oxo-norleucinyl]-6-diazo-5-oxo-norleucine against experimental tumors in conventional and nude mice

The chemotherapeutic effects of 6-diazo-5-oxo-L-norleucine (DON) and N-[N-gamma-glutamyl-6-diazo-5-oxo-norleucinyl]6-diazo-5-oxo-norleucine (azotomycin) were evaluated in a spectrum of transplantable experimental tumor systems including xenografts of human tumors in athymic mice. Both drugs displayed remarkable activity against the murine leukemia L1210 and P388, the Colon Tumors C26 and C38 and the CD8F1 mammary tumor. No significant activity was observed against Lewis lung carcinoma, B16 carcinoma, B16 melanoma, and intracranial ependymoblastoma. DON and azotomycin also exhibited striking inhibitory effects on the growth of s.c. human tumor (MX-1 mammary, LX-1 lung and CX-1 and CX-2 colon) xenografts in athymic mice. With the exception of one colon xenograft (CX-1), all tumor lines were markedly responsive to both drugs. Tumor regressions below the initial tumor sizes of 100 to 300 mg, albeit temporary, were brought about by one course of treatment every 4 days for 3 doses (at optimal dose) with either drug. Although these drugs have been tested previously in the clinic and have shown only limited therapeutic effectiveness, they seem to worthy of a second and closer look in light of the recent laboratory results.     

双β-羧乙基锗倍半氧化物对小鼠和裸鼠移植性肿瘤生长的影响

我们研究了双β-羧乙基锗倍半氧化物(Ge—132)的抗瘤作用,结果表明对小鼠Harding—Passey黑色素瘤(M—HP)、艾氏腹水癌实体瘤(ESC)和裸鼠移植人肺鳞癌的生长有明显的抑制作用,并发现Ge—132与DDP合并应用对抗裸鼠移植人肺鳞癌有协同作用。     

Lymphocyte-activating and growth-inhibitory activities for several sources of native and recombinant interleukin 1

Several native and recombinant forms of human interleukin-1 (IL-1) and recombinant murine IL-1 were assayed for their ability to inhibit the growth of cell lines established from malignant and nonmalignant human sources. The amount of growth-inhibitory activity was compared to the units of half-maximal [3H]thymidine incorporation in mouse thymocyte cultures exposed to IL-1. Three malignant human mammary cell lines (MCF-7, T47D, and MDA-MB-415) were growth inhibited in the presence of both native and the alpha and beta forms of recombinant human IL-1. MDA-MB-415 was most sensitive. Although most sources of IL-1 showed good correlation between units of activity and percentage of growth inhibition, native IL-1 from Genzyme Corporation induced a cytotoxic effect. Murine IL-1 was less growth inhibitory than the human forms of the monokine. Human embryonic lung (HEL), adult fibroblast (CRL 1445), and transformed milk (HBL-100) lines were not growth inhibited when tested against any IL-1 source. A lung carcinoma (CALU-1) and a colon carcinoma (SW-48) were not inhibited by either the alpha or beta forms of human recombinant IL-1.     

Effect of treatment schedule on antitumor activity of glycolate-0,0'-diammineplatinum(II), a new platinum derivative: comparison with cis-diamminedichloroplatinum(II)

The effect of treatment schedule on the antitumor activity of a new platinum derivative, glycolate-0,0'-diammineplatinum (II) (254S) compared with cis- diamminedichloroplatinum (II) (CDDP) was investigated using ascites L1210 leukemia and solid Lewis lung carcinoma. The drugs were given i.p. by three treatments: as a single injection, as three injections at 4 day intervals and as 9 daily continuous injections. 254S produced a marked increase of lifespan in mice by all three treatment schedules (about 100% ILS), although the consecutive treatment of 254S needed more total doses against L1210 leukemia. The antitumor activity of 254S was, however, inferior to that of CDDP. Moreover, 254S did not show certain dependence on treatment schedule, while CDDP was rather dependent on treatment schedule. The single injection (day 1) of CDDP exhibited the most potent antitumor activity. On the other hand, although the single injection of CDDP showed more host toxicity than the other treatment schedules and the consecutive treatment needed more total doses, neither drug showed any definite schedule dependency against Lewis lung carcinoma. Moreover, the tumor growth inhibitory activity of 254S was almost the same as or slightly superior to that of CDDP. Both drugs produced about 70% (days 14-17) and 50% (day 20) tumor weight inhibitions against early and advanced Lewis lung carcinoma, respectively.     

草分枝杆菌联合羟基喜树碱对SPCA/I人肺腺癌细胞抗增殖效应的研究

目的　探讨草分枝杆菌 (mycobacteriumphlei)对人肺腺癌细胞系SPCA/I的抗增殖效应 ,以及不同浓度草分枝杆菌与羟基喜树碱联合应用时的协同效应。方法　应用细胞培养和MTT比色法 ,检测不同浓度组的草分枝杆菌 ,以及草分枝杆菌与羟基喜树碱联合应用对SPCA/I人肺腺癌细胞生长的抑制作用 ,波长定于 5 70nm ,测定光密度 (A值 )并计算抑制率。结果　不同浓度的草分枝杆菌 ,在体外对SPCA/I人肺腺癌细胞株均有直接的抑制作用 ,其抑制率与所用药物浓度呈正相关。 1.72 0 μg/ml及0 .860 μg/ml浓度的草分枝杆菌与羟基喜树碱 (1.0PPC、0 .5PPC)联用 ,在体外对人肺腺癌细胞的抑制率显著高于单用羟基喜树碱组。结论　草分枝杆菌在体外对SPCA/I人肺腺癌细胞有直接抑制作用 ,该药与羟基喜树碱联合应用有显著的协同抗肿瘤效应。     

Antitumor spectra of ranimustine against various human tumors

Ranimustine (MCNU) has been shown to exhibit high antitumor activity and broad antitumor spectra against various experimental tumors. These effects were comparable to those of nimustine (ACNU). However, clinical applications of ACNU are indicated to various types of malignancies including solid tumors, while those of MCNU are almost limited to hematological ones. Therefore, the antitumor activity of MCNU was examined against 55 specimens from 15 types of solid tumors and compared with those of ACNU and 8 other drugs. Drug sensitivity was examined by a morphological method measuring the proportion of degenerative changes in the nucleus of drug-treated and untreated tumor cells. MCNU showed antitumor activities (measured by karyorrhexis) against adenocarcinoma of the lung, squamous cell carcinoma of the lung, renal cell carcinoma, bladder tumor, ovarian cancer and brain tumor. In addition, MCNU and ACNU showed a similar positive rate (15-16%) in this experiment and this was the highest among all drugs examined. Although MCNU and ACNU showed similar antitumor spectra, a clear difference was observed when the antitumor activities of both drugs were compared in each identical specimen. These results clearly suggest that MCNU is worthy of clinical study to examine the antitumor activity against various solid tumors.     

功能性单克隆抗体40E11靶向治疗肺癌的实验研究

目的:检测分析抗肺癌单克隆抗体40E11识别的靶抗原在肺癌细胞系及肺癌组织中的表达,研究单克隆抗体40E11体内外抑瘤功能,并初步鉴定其识别的抗原蛋白,为肺癌的靶向治疗提供候选抗体药物及靶标.方法:应用细胞免疫荧光检测单克隆抗体40E11所识别的靶抗原在肺癌细胞系中的表达及定位,免疫组化检测其在人肺癌组织中的表达;CCK-8法检测分析40E11对肺癌细胞增殖的影响.裸鼠体内治疗实验研究40E11对移植瘤生长的抑制作用.Western bloting鉴定该单克隆抗体识别的抗原的分子量.结果:单克隆抗体40E11识别的抗原在人肺腺癌细胞(A549、GLC-82、ANIP-973)、人肺鳞癌细胞(GLC-P)的胞内及胞膜均有表达;该抗原在84.6％的人肺癌组织中的表达较癌旁组织显著上调;CCK-8法检测结果显示单克隆抗体40E11明显抑制肺癌细胞体外生长;抗体治疗实验结果表明单克隆抗体40E11明显抑制体内肺癌细胞生长,其抑制率达64.4％,P＜0.05;蛋白质印迹法显示40E11识别的抗原相对分子质量约为100×103.结论:抗肺癌单克隆抗体40E11在体内外能显著抑制肺癌细胞的生长,为肺癌的靶向治疗提供具有重要应用价值的候选抗体药物.     

Fibrinolytic activity in human tumor tissues

The fibrinolytic activity of 156 malignant and 36 benign solid tumors from autopsy and biopsy specimens was studied by the fibrin slide technique. The inhibitory activity against fibrinolysis was graded according to the lysis time of vascular tissues within the tumor. The results show that all malignant solid tumors, with the exception of prostate carcinoma, demonstrated varying degrees of inhibition of fibrinolysis. Persistently high inhibitory activity was found in squamous cell carcinoma of the esophagus, the respiratory tract, cervix uteri, and skin; carcinoma of uterus; colorectal carcinoma; small cell anaplastic carcinoma of lung; neuroblastoma, carcinoma of bile duct, while malignant tumors of the kidney show a lesser degree of inhibition. In contrast, with the exception of the hydatidiform mole, benign solid tumors show little or no inhibition. A similar absence of fibrinolytic activity is seen in metastatic disease. Further studies of the role of the fibrinolytic system in tumors seems warranted.     

Comparative Evaluation of Antiproliferative Activity and Induction of Apoptosis by some Fluoroquinolones on a Human Non-small Cell Lung Cancer Cell Line in Culture

Lung cancer is the leading cause of cancer- related death in the world today. Since the effective management of ‍drug resistant lung cancer, and particularly non-small cell lung carcinomas is a major problem, attempts need to be ‍made to identify new potential anticancer drugs that can kill non-small cell lung cancer cells efficiently. In the ‍present study, a human non-small cell lung carcinoma NCI-H460 cell line was used to evaluate the antiproliferative ‍activity of Fluoroquinolones like Enoxacin, Norfloxacin, Ciprofloxacin and Levofloxacin. As determined by ‍Sulphorodhamine B assay (SRB assay), all Fluoroquinolones caused cellular growth inhibition in a concentration ‍and time-dependent manner. Enoxacin was found to be the most effective Fluoroquinolone followed by Norfloxacin, ‍Ciprofloxacin and Levofloxacin. Growth inhibitory effects were also found to be independent of the concentrations ‍of serum growth factors in culture medium or variation of initial cell seeding density and proved to be irreversible in ‍nature. Appearance of rounded cells with altered morphology and cell surface blebbing indicated cell killing by ‍apoptosis. Cell shrinkage, nuclear condensation & fragmentation, and cytoplasmic blebbing as indicated by MGG ‍staining confirmed this to be the case. Thus, this investigation clearly demonstrated that the NCI-H460 human nonsmall ‍cell lung carcinoma cell line is highly sensitive to Fluoroquinolone treatment. The Fluoroquinolones used in ‍this study which are clinically used as antibacterial agents, can also inhibit tumor cell growth suggesting their potential ‍use in a strategy for cancer treatment which might help in controlling cancer.     

Therapy of small cell carcinoma of the lung with hexamethylmelamine.

Hexamethylmelamine (HXM) is one of the substituted melamines derived from cyanuric chloride [1]. Previous studies with HXM have shown activity against small cell carcinoma of the lung, when it was given as a single agent [2, 3]. Because of relatively mild bone marrow toxicity, HXM was subsequently given in combination with other drugs and/or radiation therapy, in order to improve the therapeutic results. In this paper, our experience with HXM in treatment of small cell carcinoma of the lung is summarised.     

A comparison of adriamycin and mAMSA

Summary Multicellular spheroids were used to compare the two chemotherapeutic agents adriamycin (ADM) and 4[(9-acridinyl)-amino] methanesulphon-m-anisidide (mAMSA). Chinese hamster cells, V79 379A, a human small cell lung carcinoma, designated ME/MAR, and a human melanoma xenograft, HX117, were grown as spheroids (200 or 400 m in diameter) and treated with either drug for 1 h, at 37°C, in air. Cytotoxicity was assayed using both cell survival and growth delay.Both drugs were highly toxic towards V79 but showed less activity toward the human tumour single cell suspensions; ADM was more effective towards HX117 and ME/MAR than mAMSA. When grown as spheroids, the cells developed marked resistance to both drugs. In all cases, cytotoxicity was drug dose and spheroid size dependent. The response of HX117 spheroids to both drugs was similar. In contrast, ADM was more effective toward 200 m diameter ME/MAR spheroids, and mAMSA showed greater activity than ADM against V79 spheroids. Both endpoints gave qualitatively equivalent results, and a comparison of the two showed relatively long growth delays for a given level of cell kill, for both drugs and with all three cell lines. The greater cytotoxicity of ADM toward ME/MAR spheroids is consistent with the clinical finding that ADM has a use in the treatment of small cell carcinoma of the lung, while mAMSA has not demonstrated any activity in the treatment of lung cancer.     

Biological profile of FCE 24517, a novel benzoyl mustard analogue of distamycin A.

FCE 24157 (chemically (beta-[1-methyl-4-(1-methyl-4--[1-methyl-4-(4-N,N- bis(2-chloroethyl) amino-benzene-1-carboxy-amido) pyrrole-2-carboxiamido]pyrrole-2-carboxyamido)pyrrole-2-c arboxyamido]) propionamidine, hydrochloride) is a distamycin A (Dista A) derivative bearing a benzoyl mustard moiety instead of the formyl group at the N-terminal. Contrary to Dista A, FCE 24517 has been found to display potent cytotoxic activity on human and murine tumour cell lines. The compound maintains activity on melphalan (L-PAM)-resistant cells, whereas cross-resistance is observed on doxorubicin-(DX)-resistant cells. In vivo, FCE 24517 was found to possess evident antineoplastic activity on a series of murine transplanted solid tumours and human tumour xenografts. The following neoplasms were in fact found to be sensitive to FCE 24517 treatment: M14 human melanoma xenograft, N592 human small cell lung carcinoma, MTV murine mammary carcinoma, Colon 38 murine carcinoma, PO2 murine pancreatic carcinoma and M5076 murine reticulosarcoma. Lower effectiveness was observed against the murine P388 and Gross leukaemia, Lewis lung murine carcinoma, LoVo human colon carcinoma xenografts and A459 human lung adenocarcinoma. Against the murine L1210 leukaemia, FCE 24517 displayed a clear activity only when the tumour was transplanted i.p. and treatment was given i.p., whereas only marginal activity was seen against this leukaemia if transplanted i.v. and the drug was given i.v. As true also in vitro, FCE 24517 was effective against i.p. implanted L1210 leukaemia resistant to L-PAM. The mode(s) of action of this new compound is under active investigation.     

Synergistic effects of F 11782, a novel dual inhibitor of topoisomerases I and II,in combination with other anticancer agents

PURPOSE: F 11782, or 2',3'-bis-pentafluorophenoxyacetyl-4',6'-ethylidine-beta- D-glucoside of 4'-phosphate-4'-dimethylepipodophyllotoxin 2 N-methyl glucamine salt, a novel dual catalytic inhibitor of topoisomerases I and II, characterized by marked antitumour activity in vivo in a series of experimental murine and human tumours, has been selected for further development. This preclinical study was undertaken to investigate its potential for inclusion in combination chemotherapy regimens. The in vitro cytotoxicity of F 11782 incubated simultaneously with the following drugs was investigated: aclarubicin, cisplatin, doxorubicin, etoposide, 5-fluorouracil, mitomycin C, paclitaxel, topotecan or vinorelbine. METHODS: The combinations were first evaluated in vitro against the GCT27 human testicular teratoma cell line and then against the A549 human non-small cell lung cancer cell line using median effect analysis. RESULTS: F 11782 in combination with cisplatin, mitomycin C, etoposide or doxorubicin showed synergistic cytotoxicity against both cell lines. Moreover, F 11782 combined with cisplatin or mitomycin C showed antitumour activity in vivo against P388 murine leukaemia grafted intravenously. Such synergy might have resulted from the identified nucleotide excision repair inhibitory activity of F 11782. CONCLUSIONS: F 11782 appears to be a promising candidate for combination chemotherapy, especially with DNA-damaging agents.     

草分枝杆菌联合化疗药物对人肺癌细胞生长抑制作用的研究

目的　探讨草分枝杆菌对人肺腺癌细胞系SPCA/I的抗增殖作用以及不同浓度草分枝杆菌与化疗药物联合应用时的协同效应。方法　应用MTT比色法检测不同浓度组的草分枝杆菌以及草分枝杆菌与化疗药物联合应用对SPCA/I人肺腺癌细胞生长的抑制作用 ,波长定于 570nm ,测定光密度 (A值 )并计算其抑制率。结果　 (1 )不同浓度的草分枝杆菌体外对人肺腺癌细胞均有抑制作用 ,其抑制率随所用浓度呈正相关。 (2 ) 1 .72ug/ml及 0 .86ug/ml浓度的草分枝杆菌与化疗药物 (1PPC)联用 ,体外对人肺腺癌细胞的抑制率显著高于单用化疗药物组。结论　草分枝杆菌体外对SPCA/I人肺腺癌细胞有直接的抑制作用 ,该药与化疗药物联合应用有显著的协同抗肿瘤效应 ,其结果可能为肿瘤的临床联合化疗提供实验依据     

异种免疫诱导中和性抗体抑制小鼠肺癌生长

目的 研究异种固定化完整细胞作为肿瘤疫苗的潜力,初步探讨其诱导的免疫反应的抗肿瘤机制。方法 分别使用固定的人肺癌细胞免疫小鼠,再接种小鼠自发肺腺癌鼠肿瘤进行肿瘤挑战,观察肿瘤细胞免疫后对小鼠肿瘤生长的影响。采用免疫组织化学法观察小鼠肿瘤浸润T细胞的情况,采用MTT法检测小鼠免疫血清对人肺癌细胞生长的影响。结果 采用人肺腺癌和鳞癌固定的完整细胞免疫小鼠后均明显地抑制小鼠肺腺癌肿瘤的生长（P<0.05）,抑瘤率分别为40.8%及49.2%,显著优于采用人肺腺癌和鳞癌细胞碎片免疫的效果。小鼠免疫血清在体外可抑制人肺癌细胞的生长。结论 异种人肺癌固定的完整细胞诱导了有效的小鼠抗鼠肺癌的体液免疫反应,显著抑制肿瘤生长,其抑制作用可能与诱导小鼠产生抗肺癌中和性抗体有关。     

Antitumor activity of a new platinum compound (glycolate-o,o') diammineplatinum (II) (254-S), against non-small cell lung carcinoma grown in a human tumor clonogenic assay system

(Glycolate-o,o') diammineplatinum (II) (254-S) is one of the platinum derivatives showing high activity against rodent solid tumors and lack of renal toxicity. We have used a human tumor clonogenic assay (HTCA) as a disease-oriented drug screening model for new antitumor drugs, in order to test the antitumor activity of 254-S against non-small cell lung carcinoma (NSCLC) and to compare its activity with that of cisplatin (CDDP) and of carboplatin (CBDCA). The overall in vitro response rate (defined as less than 50% survival of tumor colony forming units) for 254-S was observed in 10/29 and 20/30 evaluable specimens isolated freshly from NSCLC patients with continuous exposure at 1 and 10 micrograms/ml, respectively, indicating a positive dose-response relationship. Dose dependent cytotoxicity was also confirmed in 4 human tumor cell lines derived from NSCLC patients. The antitumor activity of 254-S was 3.6-fold that of CBDCA, but two-fifths that of CDDP. A comparison of these in vitro results with the toxic properties of 254-S such as low nephrotoxicity suggests that 254-S is a promising new drug against NSCLC.     

伊立替康、奥沙利铂及氟尿嘧啶对人大肠癌LoVo细胞株作用的实验研究

目的：研究国产盐酸伊立替康（CPT-11）、奥沙利铂（L-OHP）及氟尿嘧啶（5-FU）对人大肠癌细胞株LoVo的生长抑制作用。方法：采用MTT法观察上述药物单药应用、两药及三药同时或序贯联合应用对大肠癌LoVo细胞生长的影响，评价药物联合应用的效果。结果：单药应用、两药联合及三药联合应用均对大肠癌LoVo细胞有显著的抑制作用（P〈0．05）；两药联合应用均优于各自单药的抑制效果，CPT-11联合L-OHP弱于CPT-11或L-OHP与5-FU联合的抑制效果（P〈0．05），由L-OHP到CPT-11小剂量序贯联合具有明显的协同作用；三药联合应用的抑制效果显著优于两药联合（P〈0．05），由5-FU＋L-OHP到5-FU＋CPT-11的序贯联合应用与三药同步联合应用无显著性差异（P〉0．05），但由5-FU＋CPT-11到5-FU＋L-OHP序贯应用时明显低于三药同步应用的效果（P〈0．05），且三药全量及半量联合应用效果差异无显著性（P〉0．05）。结论：由5-FU＋L-OHP到5-FU＋CPT-11的小剂量序贯联合应用具有高效协同抑制大肠癌细胞株LoVo的作用，可为临床难治性和复发性大肠癌患者的化疗及小剂量序贯化疗提供参考。     

Human tumor-cell growth modulatory effects of the aebs/hic-binding drugs used as single agents and in combination with a novel amphibian oocyte rnase

A novel amphibian oocyte RNase, ONCONASE(R) (ONC), has been previously shown to have synergistic tumor cell growth inhibitory activity when combined with tamoxifen (TMX) and/or trifluoperazine (TFP) in human ASPC-1 pancreatic and A-549 lung carcinoma cells, respectively. It has recently been reported that several drugs known to bind to the intracellular antiestrogen binding site (AEBS)/histamine (H(IC)) receptors, including tricyclic (amitriptyline, AMT) and non-tricyclic (fluoxetine, FLX) antidepressants, TMX, phenothiazines and the prototype H(IC)-binder DPPE, can stimulate the in vitro and in vivo growth of rodent tumor cells, while having a normal cell growth inhibitory activity, as reflected by the inhibition of the DNA synthesis. It has been presently shown that while at the clinically relevant concentrations some of these H(IC)-binding drugs mildly stimulated (up to 15%) the cell growth in the human lines studied when used as single agents, in most instances this stimulation did not exceed 10% above the control values. When used in combination with ONC, neither of these H(IC)-binding drugs demonstrated any apparent synergistic activity as judged from the ED50 values. However, the combinations of DPPE+TFP and AMT+TFP, in both the ASPC-1 and COLO 320DM lines, demonstrated a significant cell growth inhibition, while there was no difference between the effects of AMT alone and the AMT+TFP combination in the U87MG line. The most effective cell growth inhibition was obtained when ONC was combined with DPPE+TFP and/or AMT+TFP, as reflected by the significantly decreased ED50 values.     

A new in vitro screening system for anticancer drugs for the treatment of non-small cell lung cancer

We have evaluated a semiautomated radiometric assay (BACTEC 460 system) for screening of activity of anticancer drugs against human non-small cell lung cancer cell lines. Cells from seven cell lines were exposed to standard antineoplastic agents at four different concentrations using a 1-h incubation. Alpha 2-interferon was tested using a continuous incubation. In vitro drug activity was analyzed as a function of the clinically achievable serum concentration. Our results indicate that two cell lines (CALU-3, SK-MES-1) exhibit in vitro drug sensitivity patterns closest to those observed in clinical studies. These two cell lines might therefore be most useful for screening new anticancer compounds for activity against non-small cell lung cancer. The radiometric assay is a semiautomated system which has advantages over other, more time-consuming screening systems.