草分枝杆菌对SPCA/Ⅰ人肺癌细胞生长抑制作用的研究

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.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.结果不同浓度的草分枝杆菌体外对人肺腺癌细胞株SPCA/Ⅰ均有直接的抑制作用,其抑制率与所用药物浓度呈正相关.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,该结果可能为肿瘤的临床联合治疗提供实验依据.     

草分枝杆菌对SPCA／I人肺癌细胞生长抑制作用的研究

目的：探讨草分枝杆菌（Mycobacterium Phlei)对人肺腺癌细胞系SPCA／I的抗增殖效应。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：不同浓度的草分枝杆菌体外对人肺腺癌细胞株SPCA／I均有直接的抑制作用,其抑制率与所用药物 浓度呈正相关。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,该结果可能为肿瘤的临床联合治疗提供实验依据。     

草分枝杆菌调节SPCA/Ⅰ人肺腺癌细胞生长周期的研究

目的探讨草分子杆菌对SPCA/Ⅰ人肺腺癌细胞系的抗增殖效应及调节细胞生长周期的作用。方法将SP-CA/Ⅰ人肺腺癌细胞株接种于细胞培养液(RPMI-1640)培养传代,分别加入不同浓度的草分子杆菌,作用36小后,分别以酶标仪检测细胞抑制率,倒置显微镜观察凋亡细胞形态变化,以cycletest plus DNA reagent kit标记,经FACSAria流式细胞仪检测细胞周期改变。结果①不同浓度的草分子杆菌体外对人肺腺癌细胞株SPCA/Ⅰ均有直接的抑制作用,其抑制率与所用药物浓度呈正相关。②倒置显微镜下可见细胞由原来贴壁生长而逐渐脱落、外形变圆、体积缩小、折光性增强,核染色质凝集,细胞裂解。③草分子杆菌体外能改变SPCA/Ⅰ人肺腺癌细胞株的细胞增殖周期,使细胞周期阻滞在G2~M期,细胞凋亡率明显增加。结论草分子杆菌能通过调节细胞增殖周期、诱导细胞凋亡,直接抑制人肺腺癌细胞株SPCA/Ⅰ增殖。     

三氧化二砷对SPCAⅠ/人肺腺癌细胞株抗增殖及诱导凋亡作用的研究

目的探讨三氧化二砷(As2O3)对SPCAⅠ/人肺腺癌细胞株的抗增殖效应及诱导细胞凋亡的作用。方法将SPCAⅠ/人肺腺癌细胞株培养传代,分别加入不同浓度的三氧化二砷,作用24小时后,分别以酶标仪检测细胞抑制率、FACSAr-ia流式细胞仪检测细胞凋亡率、倒置显微镜观察凋亡细胞形态。结果不同浓度的三氧化二砷体外对人肺腺癌细胞株SPCA/Ⅰ均有直接的抑制作用,其抑制率与所用药物浓度呈正相关;三氧化二砷体外对人肺腺癌细胞株SPCAⅠ/有诱导凋亡的作用,细胞凋亡率明显增加。倒置显微镜见细胞由原来贴壁生长而逐渐脱落、外形变圆、体积缩小、折光性增强,核染色质凝集,细胞裂解。结论三氧化二砷能通过诱导细胞凋亡,直接抑制人肺腺癌细胞株SPCAⅠ/增殖。     

三氧化二砷对SPCA/I人肺腺癌细胞株抗增殖及诱导凋亡作用的研究

目的 探讨三氧化二砷( As2 O3)对SPCA/I人肺腺癌细胞株的抗增殖效应及诱导细胞凋亡的作用.方法 将SPCA/I人肺腺癌细胞株培养传代,分别加入不同浓度的三氧化二砷,作用24小时后,分别以酶标仪检测细胞抑制率、FACSAria流式细胞仪检测细胞凋亡率、倒置显微镜观察凋亡细胞形态.结果 不同浓度的三氧化二砷体外对人肺腺癌细胞株SPCA/I均有直接的抑制作用,其抑制率与所用药物浓度呈正相关;二氧化二砷体外对人肺腺癌细胞株SPCA/I有诱导凋亡的作用,细胞凋亡率明显增加.倒置显微镜见细胞由原来贴壁生长而逐渐脱落.外形变圆、体积缩小、折光性增强,核染色质凝集,细胞裂解.结论 三氧化二砷能通过诱导细胞凋亡,直接抑制人肺腺癌细胞株SPCA/I增殖.     

黄芪对肺腺癌细胞生长抑制的实验研究

目的:研究分化诱导剂黄芪(Astragalus)在体外对人肺腺癌SPC-A-1细胞生长抑制的影响。方法:用不同浓度的黄芪分别处理肺腺癌SPC-A-1细胞后,镜下观察癌细胞的生长情况;测定软琼脂克隆形成率;MTT(噻唑蓝)法测定生长抑制率。结果:不同浓度黄芪处理后细胞的生长明显受到抑制,细胞生长抑制率随黄芪的浓度增加而增加。结论:黄芪对肺腺癌SPC-A-1细胞的生长有明显的抑制作用。     

香叶木素对人肺腺癌细胞系H1299和A549增殖能力影响的研究

目的:探究香叶木素(Dios)作用前后人肺腺癌细胞系H1299和A549细胞增殖活性的改变。方法:取RPMI-1640培养基培养的人肺腺癌细系H1299及A549用于实验。采用CCK8法检测Dios作用前后人肺腺癌细胞系H1299和A549细胞增殖活性的改变。结果:不同浓度(0、5、10、20μmol/L)Dios作用H1299细胞24h后,细胞的增殖抑制率分别为(10.9±2.5)%、(32.7±1.8)%、(41.2±2.1)%、(51.7±2.8)%,而A549细胞增殖抑制率为(18.9±2.3)%、(30.8±2.6)%、(42.2±1.2)%、(50.7±2.1)%。结论:Dios对人肺腺癌细胞系H1299和A549的增殖有明显的抑制作用,提示Dios通过抑制肺腺癌细胞的增殖,进而起到良好的抗肿瘤作用。     

红景天对体外培养肺腺癌细胞增殖的抑制作用

目的研究红景天(Rhodiola)对体外培养肺腺癌SPC-A-1细胞的直接抑制作用及可能机制。方法通过液体培养法、克隆形成法、3H-TdR掺入法与MTT比色法分别检测给药后肺腺癌细胞的增殖情况、克隆形成率、DNA合成抑制率及生存率来探讨红景天的抑瘤效应。结果与红景天共育24小时的细胞伸展不良,胞体回缩,贴附型细胞不贴壁,胞质粗糙,有大量颗粒状物堆积,而且药物浓度越大,形态学改变越明显。给药组肿瘤细胞增殖缓慢甚至停滞,出现细胞脱落、胞浆内颗粒状物堆积等形态学改变,克隆形成数明显少于对照组,3H-TdR掺入率减少,生存率下降。结论红景天对体外培养肺腺癌细胞均具有直接杀伤作用。红景天可能通过干扰细胞代谢、改变细胞外衣的性质抑制肿瘤细胞增殖。     

昆明山海棠对肺腺癌细胞株生长抑制观察

目的：探讨昆明山海棠对培养人肺腺癌细胞株的抗癌效果。方法：采用贴壁细胞培养盖玻片法及MTT显色检测药物对癌细胞生长抑制实验。结果：10微克／毫升昆明山海棠水煎剂对癌细胞生长抑制率二种指标均达到70％。结论：昆明山海棠水煎剂在体外对培养人肺腺癌细胞有明显的细胞毒作用。     

姜黄素对人SPC-A549细胞生长抑制的影响

目的　研究姜黄素对人肺腺癌细胞SPC -A5 4 9的生长抑制作用。方法　采用细胞培养 ,倒置相差显微镜下观察细胞形态变化 ;MTT法检测姜黄素对肺腺癌细胞的抑制作用。结果　①姜黄素作用癌细胞后 ,光镜下可见细胞脱壁 ,胞体缩小 ,胞核固缩。② 5、10、2 0和 4 0mg/L姜黄素均能抑制人肺腺癌细胞SPC -A5 4 9的生长 ,4 0mg/L姜黄素作用 12h细胞存活率达2 8.5 %。结论　姜黄素能有效的抑制人肺腺癌细胞SPC -A5 4 9的生长 ,且对人SPC -A5 4 9细胞存活率具有浓度和时间依赖性     

肿瘤坏死因子与化疗药物协同抑制肺癌细胞生长的研究

目的：探讨肿瘤坏死因子α对人肺腺癌细胞系A549的毒性作用及与化疗药物联合应用的协同效应，以期为肺癌的临床治疗提供新的依据。方法：应用MTT显色法检测不同浓度组药物对A549细胞的毒性作用，并计算其抑制率。结果：肿瘤坏死因子α能直接抑制人肺腺癌A549细胞的生长,嘬大抑制率为23．06％。低剂量的肿瘤坏死因子（100U／ml，50U／ml)与化疗药物（IPPC、0．1KPPC）联用后的抑制率高于单用化疗药物，其中以与MMC、VCRK、5－FU联用后的效果最好，最大抑制率均在80％左右。结论：（1）肿瘤坏死因子α对A549细胞具有直接抑制效应，但敏感性不高、肿瘤坏死因子α与部分化疗药物联合应用对A549细胞系具有显著的协同抑制效应。这些可能为肺癌的临床治疗提供实验依据。     

EFFECTS OF p53 GENE THERAPY COMBINED WITH CYCLOOXYGENASE-2 INHIBITOR ON CYCLOOXYGENASE-2 GENE EXPRESSION AND GROWTH INHIBITION OF HUMAN LUNG CANCER CELLS

1 INTRODUCTIONCOXis the key enzyme involved in the synthesis of prostanoids,a collective termfor the PGs andthromboxanes.Of the two major isoforms of the COXenzyme,COX-1 is ubiquitous and constitutively ex-pressedin virtually all normal tissues.In contras…     

Endostatin exhibits a direct antitumor effect in addition to its antiangiogenic activity in colon cancer cells

Endostatin has been considered a highly specific inhibitor of endothelial cell proliferation and/or migration. To explore the use of endostatin in antiangiogenic gene therapy, we generated a recombinant adenovirus, AdEndo, carrying the gene for mouse endostatin. Injection of 10(9) PFU of AdEndo resulted in a low but significant suppression (25%) of preestablished tumor growth in murine models involving murine Lewis lung carcinoma (LLC) and human breast cancer MDA-MB-231 tumors. Greater anticancer activity was observed when the same dose of AdEndo was injected into two other preestablished murine models involving C51 murine colon cancer and HT29 human colon cancer (55 and 47% tumor growth reduction, respectively). In vitro, endostatin derived from AdEndo-infected MRC-5 fibroblasts inhibited the growth of C51 and HT29 cell lines (72 and 61%, respectively). The extent of this inhibition was comparable to that observed in endothelial cells: 75% for microcapillary endothelial cell line HMEC-1, 52% for human dermal microvascular endothelial cells, 46% for human umbilical vein endothelial cells, and 67% for calf pulmonary arterial endothelial cells. Both endothelial and colon cancer cells showed a clear increase in cell apoptosis (4- to 5-fold for endothelial cells and 5- to 10-fold for colon cancer cells) and an accumulation in the G(1) phase of the cell cycle. This antiproliferative activity was not observed in other tumor cell lines: LLC, MDA-MB-231, murine colon adenocarcinoma MC38, human prostate cancer cell line DU145, and human breast cancer cell line CAL51. Taken together, these results provide evidence that, in addition to its antiangiogenic activity, endostatin exerts a direct anticancer action that appears to be restricted to some tumor cell lines. Thus, endostatin could be used in some colon cancer treatments and its clinical efficacy would depend on the response of tumor cells themselves.     

双氢青蒿素对人肺腺癌细胞株A549抑制作用的实验研究

目的观察双氢青蒿素对人肺腺癌细胞株A549的作用。方法采用四唑氮蓝（MTT）法测定不同浓度和时间双氢青蒿素对A549细胞的生长抑制作用;应用流式细胞术检测双氢青蒿素对A549细胞增殖和细胞周期的影响;应用透射电镜分析法检测双氢青蒿素对A549细胞凋亡的影响。结果双氢青蒿素对A549细胞有明显的体外增殖抑制作用,72 h的IC50值为580 nmol/L,且有明显的时间和剂量依赖关系;双氢青蒿素作用后G0/G1期细胞数增加;双氢青蒿素作用A549细胞后可以出现凋亡的形态学改变。结论双氢青蒿素对人肺腺癌A549细胞有生长抑制作用,其机制可能与诱导A549细胞周期阻滞和凋亡有关。     

The effects of combined treatment with sevoflurane and cisplatin on growth and invasion of human adenocarcinoma cell line A549

Sevoflurane, an inhalational anesthetic, and cisplatin (DDP)-based chemotherapy have been widely used during lung cancer surgery. However, the effect of sevoflurane on the sensitivity of lung cancer cells to DDP chemotherapy remains unclear. In this study, the effects of combined treatment with sevoflurane and cisplatin on the growth and invasion of human lung adenocarcinoma A549 cell line have been investigated. The underlying mechanism has also been explored. In our experiment, A549 cells were treated with 2.5% sevoflurane, 10 μmol/L DDP, or the co-treatment of sevoflurane and DDP for 4 h, respectively. Cell proliferation was evaluated by the MTT assay and colony formation assay. Apoptosis was assessed by flow cytometry. Cell invasion was detected by Transwell assay. The expressions of X-linked inhibitor of apoptosis protein (XIAP), Survivin, matrix metalloproteinase (MMP)-2 and MMP-9 were determined by western blotting. Our results showed that sevoflurane combined with DDP resulted in a more pronounced inhibition of tumor cells growth and invasion as compared with either drug alone. Besides, XIAP, Survivin, MMP-2, and MMP-9 were downregulated more significantly by the co-treatment of the two drugs as compared to sevoflurane treatment or DDP treatment alone. Taken together, the growth-inhibitory and invasion-inhibitory synergy between sevoflurane and DDP in human adenocarcinoma A549 cell line was found in this study. Furthermore, we showed that the growth-inhibitory synergy between sevoflurane and DDP might be associated with the downregulation of XIAP and Survivin, and the invasion-inhibitory synergy between sevoflurane and DDP might be involved in the downregulation of MMP-2 and MMP-9.     

Cell cycle arrest by oxaliplatin on cancer cells

Oxaliplatin (L-OHP) is the only platinum compound to show activity in colorectal cancer. We evaluated the cytotoxicity of L-OHP on four human cancer cell lines and its influence on the cell cycle, when treated during long exposure (72 h) and different post-incubation times (24 or 72 h). We used a panel of cell lines: HT29 (colon cancer), MCF7 (breast cancer), Hela (uterine cervix) and A549 (lung adenocarcinoma). Inhibition concentration (IC)(50) was assessed by MTT assay. Cell cycle modifications were determined using dual parameter bromodeoxyuridine and propidium iodide. L-OHP yielded a superior cytotoxicity on HT29 and MCF7 relative to Hela and A549 after treatment, the post-incubations demonstrate that growth inhibition was irreversible for HT29 and Hela cell lines contrary to MCF7 and A549. The main effects of L-OHP are G2/M cell cycle arrest and transient S phase delay. Taken together, L-OHP treatment results on HT29, MCF7 and Hela, are in favor of lengthening the infusion duration to patients during further clinical trials.     

The effects of cetuximab alone and in combination with endostatin on vascular endothelial growth factor and interleukin-8 expression in human lung adenocarcinoma cells

Background: Angiogenesis, the growth of new blood vessels, plays an important role in tumor growth and metastasis. Both cetuximab and endostatin have been found to reduce the expression of endothelial-stimulating growth factors such as vascular endothelial growth factor (VEGF) and interleukin (IL)-8. However, the effects of cetuximab alone or in combination with endostatin on human lung adenocarcinoma cell growth remain unclear.Objective: The aim of this study was to evaluate the cellular and molecular effects of cetuximab alone and in combination with endostatin on human lung adenocarcinoma cell lines HI 299, SPC-A1, and H460 in vitro.Methods The epidermal growth factor receptor (EGFR) status of a panel of human lung adenocarcinoma cell lines was characterized using Western blot analysis. We used a modified tetrazolium salt assay to evaluate the growth-inhibitory effects of cetuximab and endostatin alone and in combination on the cell lines. We also determined the effects of these 2 drugs on VEGF and IL-8 expression using enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Cells were treated for 4 days with cetuximab 12.5 μ/mL, endostatin 25 μ/mL, or cetuximab 12.5 μg/mL + endostatin 25 μg/mL. Untreated cells cultured for 4 days served as controls.Results: EGFR expression in the H1299 cells was higher than in the SPC-A1 and H460 cells. Varying concentrations of cetuximab alone were associated with a significant growth-inhibitory effect on all 3 cell lines in a dose-dependent manner after 4 days of exposure compared with controls (all, P < 0.05). Compared with controls, varying concentrations of endostatin alone were not associated with significant inhibition of cell growth in any of the 3 cell lines. The inhibitory ratio of cetuximab + endostatin at varying concentrations was significantly greater than that of cetuximab alone (all, P < 0.05). On ELISA, either drug alone was associated with significant reductions in secreted VEGF and IL-8 in the HI 299, SPC-A1, and H460 cell lines (all, P < 0.05), with the exception of IL-8 concentration in the H460 cells. Mean (SD) VEGF expression with combination treatment in the H1299 and SPC-A1 cell lines (687 [21] and 629 [23] pg/mL, respectively) was significantly lower than with cetuxi-mab alone (878 [31] and 708 [20] pg/mL; both, P < 0.001); in the H460 cell line, combination treatment was not associated with a significant further reduction in VEGF expression. IL-8 concentrations with cetuximab in the H1299, SPC-A1, and H460 cell lines were 628 (20), 484 (29), and 532 (28) pg/mL, respectively, while the IL-8 concentrations with the combination treatment were 516 (20), 480 (18), and 467 (30) pg/mL. An enhanced effect of endostatin on IL-8 was observed in the H1299 and H460 cell lines (P < 0.001 and P = 0.018, respectively); however, no enhanced effect in the SPC-A1 line was observed. Similar results for VEGF and IL-8 expression were found using Western blot analysis.Conclusions: The results from this in vitro study suggest that cetuximab treatment might both inhibit human lung adenocarcinoma cell line growth and reduce the expression of VEGF and IL-8, which are the biomarkers of angiogenesis. Endostatin was not associated with inhibition of human lung adenocarcinoma cell line growth directly. Findings with the combination of cetuximab + endostatin suggest that endostatin might enhance the antiangiogenic and antitumor activity of cetuximab through an apparent effect on VEGF expression and, to a lesser degree, on IL-8 expression.