加温诱导肿瘤细胞凋亡与逆转多药耐药性的实验研究

目的　探讨加温逆转肿瘤细胞株ＫＢＶ２００ 多药耐药性及与诱导细胞凋亡的关系。方法用四唑蓝快速比色法（ＭＴＴ法） 检测细胞存活率;用形态学（ 光镜、电镜观察） ;生化学（ＤＮＡ 末端转移标记法,ＴＵＮＥＬ法） 及流式细胞术分析细胞凋亡状况。结果　加温４２ ℃,１ ｈ 可明显降低ＫＢＶ２００ 细胞存活率,提高长春新碱细胞毒性作用;单加ＶＣＲ 细胞存活率为（９０ ．７０ ±８．７８） ％ ,加ＶＣＲ 加温为（５７ ．４０±２１ ．５５）％ ,Ｐ＜ ０ ．００２。加温与细胞分化剂ＣＤＡ－ Ⅱ合用,有明显协同逆转作用。ＣＤＡ－ Ⅱ与ＶＣＲ合用细胞存活率为（５９ ．２４ ±３０．８０）％ ,加温与ＣＤＡ－ Ⅱ及ＶＣＲ 合用为（１５ ．３２ ±２ ．６８）％ ,Ｐ＜ ０．０５。４２ ℃或４３ ℃处理１,２４ ｈ 后即可观察到明显细胞凋亡。４３ ℃１ ｈ 处理诱导作用更强。结论　加温可明显逆转肿瘤细胞多药耐药及诱导细胞凋亡。加温对肿瘤细胞凋亡的诱导,在加温逆转肿瘤细胞耐药性中可能起着重要作用。     

Chemotherapeutic Activities of Carthami Flos and Its Reversal Effect on Multidrug Resistance in Cancer Cells

Multidrug-resistance (MDR) represents a major cause of failure in cancer chemotherapy. The need for a reduction in MDR by natural-product-based drugs of low toxicity led to the current investigation of applying medicinal herbs in future cancer adjuvant therapy. Carthami Flos (CF), the dried flower of safflower (Carthamus tinctorius L.), is one of the most popular traditional Chinese medicinal herbs used to alleviate pain, increase circulation, and reduce blood-stasis syndrome. The drug resistance index of the total extract of CF in MDR KB-V1 cells and its synergistic effects with other chemotherapeutic agents were studied. SRB cell viability assays were used to quantify growth inhibition after exposure to single drug and in combinations with other chemotherapeutic agents using the median effect principle. The combination indexes were then calculated according to the classic isobologram equation. The results revealed that CF showed a drug resistance index of 0.096. In combination with other chemotherapeutic agents, it enhanced their chemo-sensitivities by 2.8 to 4.0 folds and gave a general synergism in cytotoxic effect. These results indicate that CF could be a potential alternative adjuvant antitumour herbal medicine representing a promising approach to the treatment of some malignant and MDR cancers in the future.     

核因子-κB活性在创伤性炎症大鼠肝脏损伤中的变化及其意义

目的探讨核因子-κB(NF-κB)活性在创伤性炎症大鼠肝脏损伤中的变化及其意义。方法 Wistar 大鼠60只,随机分成对照组和创伤性炎症组。运用匀浆法提取肝细胞的核蛋白,运用凝胶阻滞实验检测创伤性炎症术后肝脏组织NF-κB的活性,检测血浆转氨酶的水平,在光镜下观察肝细胞损伤程度,电镜下观察肝脏超微结构改变,以线粒体的肿胀程度作为肝细胞损伤的评价指标。结果大鼠创伤性炎症术后第3小时,肝脏NF- κB的活性开始升高,第12小时达高峰,第72小时后基本降至正常。血浆ALT含量在伤后明显上升,于第24小时达高峰,同时肝细胞出现明显的水肿、变性和坏死,肝小叶结构破坏明显。结论大鼠创伤性炎症肝脏损伤后,NF-κB活性明显升高,与肝脏结构和功能改变基本一致,NF-κB在创伤性炎症肝脏损伤中具有重要意义。     

Long-lasting Accumulation of Vinblastine in Inostamycin-treated Multidrug-resistant KB Cells

Inostamycin, a novel polyether compound, reverses multidrug resistance in KB cells. The mechanism of its action was studied by use of radioactively labeled vinblastine. Inostamycin dose-dependently increased the accumulation of [³H] vinblastine in multidrug-resistant KB-C4 cells at 0.5-2 μg/ml, while it did not enhance accumulation in the drug-sensitive KB-3-1 cells. At a concentration of 1 μ/ ml inostamycin inhibited active [³H] vinblastine efflux from KB-C4 cells, but not from KB-3-1 cells, and inhibited [³H] vinblastine binding to KB-C4 membranes with an IC_5O of 0.94 μg/ml (1.3 μ M ). Furthermore, [³H] vinblastine accumulated by treatment with 1 /μg/ml of inostamycin was resistant to efflux from KB-C4 cells, even after the removal of inostamycin.     

Higher incidences of anti-SS-A/Ro and anti-SS-B/La autoantibodies in Japanese patients with autoimmune disorders —studies of antigens and antibodies using the immunoblotting method

Summary A specific and sensitive assay was performed to detect both anti-SS-A/Ro, and anti-SS-B/La antibodies in sera of patients with autoimmune disease, including systemic lupus erythematosus, (SLE), progressive systemic sclerosis (PSS), Sjögren's syndrome (SS), discoid lupus erythematosus (DLE), mixed connective tissue disease (MCTD), generalized morphea (GM), and dermatomyositis (DM). The SS-A/Ro and SS-B/La antigens were prepared from human spleen (HSE) and cultured human cell line (KB cells, KBE), while rabbit thymus extract (RTE) was used as the SS-B/La antigen marker. The antigens were partially purified by DEAE cellulose column chromatography. Immunoblotting showed that the SS-A/Ro antibody reacts mainly with the 58-kDa peptide of the partially purified antigen. Sera containing both the SS-A/Ro and SS-B/La antibodies reacted with the 40-kDa peptide of RTE, and the 58-kDA, 42-kDa, and 40-kDa peptides of HSE and KBE. We found that some of the SS-A/Ro antisera could further react with the 64-kDa peptide of HSE and KBE. The 58-kDa peptide is rich in its cytoplasmic fraction of KB cells, and the 40-kDa peptide in nucleoplasmic fraction. The KB cell line is a better source of the antigens than human spleen extract. The immunoblotting method clearly showed that the positivity rates of SS-A/Ro and/or SS-B/La auto-antibodies were higher in sera from Japanese patients with SLE compared with titers reported for Caucasians but not in sera from healthy volunteers.     

纳米化紫杉醇与电离辐射对KB细胞联合作用的研究

目的研究纳米化紫杉醇（TAX-NLC）与电离辐射对KB细胞杀伤的联合作用。方法利用MTT法观察TAX-NLC、电离辐射及其二者联合对KB细胞的杀伤效应。通过流式细胞仪观察TAX-NLC及电离辐射对细胞周期的影响。结果 TAX-NLC较紫杉醇（TAX）对KB细胞具有更强杀伤作用,TAX-NLC或TAX与电离辐射联合作用时,随着剂量的加大,对KB细胞的杀伤作用增强。与同等剂量的电离辐射联合作用时,同浓度的TAX-NLC也比TAX的作用强。同样,TAX-NLC可以影响细胞周期再分布,使G_2/M期细胞比例增加,与电离辐射联合作用时,其对细胞周期的影响要强于TAX。结论 TAX-NLC对KB细胞的杀伤作用高于TAX。TAX-NLC与电离辐射的联合效应比与TAX的联合效应要高。TAX-NLC作用机制可能是其更易通过细胞膜并在细胞中浓聚有关。     

Prognostic Value of Phosphorylated mTOR/RPS6KB1 in Nonsmall Cell Lung Cancer

Background: The mammalian target of rapamycin (mTOR) /RPS6KB1 activation has recently been implicatedin tumour development, but its role in lung cancer remains unclear. The aim of this study was to explore the roleof mTOR/RPS6KB1 signaling pathway in non-small-cell lung cancer (NSCLC). Methods: Immunohistochemistrywas performed to assess the expression of phosphorylated mammalian target of rapamycin (p-mTOR) and itsdownstream ribosomal phosphorylated RPS6KB1 (p-RPS6KB1) in NSCLC patients. We also analyzed p-mTOR/p-RPS6KB1 protein expression in 45 fresh NSCLC tissues using Western blotting. Results: The expressionlevel of p-mTOR and p-RPS6KB1 was significantly higher in NSCLC tumor specimens than that in adjacentnoncancerous normal lung tissues (P<0.01). p-mTOR expression correlated with p-RPS6KB1. Furthermore,high expression level of p-mTOR or p-RPS6KB1 in NSCLC was associated with a shorter overall survival (bothP<0.01). Multivariate analysis indicated high level of p-mTOR expression was an independent prognostic factor(HR=2.642, 95%CI 1.157–4.904, p=0.002). Conclusions: p-mTOR and p-RPS6KB1 could be useful prognosticmarkers for NSCLC.