甘氨双唑钠对Tca-8113细胞体外放射增敏及黏附作用的影响

背景与目的:舌鳞状细胞癌是口腔癌中发病率较高的一种,治疗方法以手术加放疗为主.然而放疗常因射线对正常组织损伤或肿瘤对射线的抵抗而受到影响.因此,选择良好的放射增敏剂已成为优化舌癌治疗的新途径.本研究旨在探讨甘氨双唑钠(glycididazole sodium for injection,CMNa)联合射线对舌鳞状细胞癌Tca-8113细胞放射增敏及黏附的影响.方法:取对数生长期的Tca-8113细胞作为研究对象,MTT比色实验检测CMNa对Tca-8113细胞增殖的影响.克隆形成实验检测CMNa联合不同放射剂量对Tca-8113细胞克隆形成能力的影响,用多靶单击模型拟合细胞生存曲线,得出放射生物学敏感参数Do(平均致死剂量)、Dq(准阈剂量),并计算放射增敏比.流式细胞术FITC和PI双染比较不同放射剂量与CMNa联合不同放射剂量射线处理后细胞凋亡的变化.同质黏附实验检测CMNa联合放射处理前后细胞黏附能力的改变.结果:Tca-8113细胞的存活率随CMNa作用浓度及作用时间的延长而降低.克隆形成实验中求得单纯放射组的Do值为2.50,CMNa联合放射线处理的Do值为1.48,放射增敏比为1.68.流式细胞术检测CMNa联合不同放射剂量(2、4和6 Gy)的细胞凋亡率(49.6%、63.1%和78.8%)均显著高于单纯放射组(30.0%、53.9%和61.2%,P＜0.01).同质黏附实验在振荡20 min时,单纯放射组及CMNa+放射组与对照组相比,黏附细胞数显著增高(P＜0.01);但单纯放射组与CMNa+放射组相比黏附细胞数差异无统计学意义(P＞0.05).在振荡40和60 min时,单纯放射组及CMNa+放射组与对照组相比,黏附细胞数显著增高(P＜0.01);且(CMNa+放射组与单纯放射组相比,黏附细胞数差异有统计学意义(P＜0.01).结论:CMNa对Tca-8113细胞有明显的放射增敏作用,其机制可能与促进细胞凋亡有关,并可能改变Tca-8113细胞的同质黏附能力.     

PsD007对四株NPC细胞放射敏感性的影响

采用ＭＴＴ方法观察癌光啉结合＾６０Ｃｏγ射线放射对四株人鼻咽癌细胞体外放射增敏作用，结果表明在一定的ＰｓＤ００７浓度范围内，随ＰｓＤ００７浓度的升高，放射对细胞的杀灭作用明显增强，当浓度分别为１．２５，６．２５，１２．５，２５和５０μｇ／ｍｌ时，对四株细胞杀灭增加倍数分别为１．０１－１．０６，１．１５－１．２９，１．２８－１．５９，１．７９－３．１８，２．８６－５．５５；放射量效存活曲线按多靶单击     

非小细胞肺癌放疗敏感性miRNAs的生物信息学筛选

目的利用基因芯片及生物信息学方法预测与非小细胞肺癌(NSCLC)放疗敏感性相关的miRNAs。方法检索GEO数据库,获取H460、H1299细胞株经2Gy电离辐射后随时间变化的基因表达谱,经组织特异性分析、功能富集(GO)分析、信号转导通路(Pathway)分析、miRNAs预测分析、转录因子(TF)分析,并建立mRNA-miRNA-TF网络,获取NSCLC放疗敏感性相关的miRNAs。结果获得了NSCLC放疗敏感性基因表达谱,辐射抵抗潜在基因共23个,辐射敏感潜在基因共97个;与放疗敏感性相关的miRNAs共19个,其中调控辐射抵抗组的miRNAs共14个:hsa-miR-1-3p、hsa-miR-1297、hsa-miR-153-3p、hsa-miR-193a-3p、hsa-miR-206、hsa-miR-302e、hsa-miR-328-3p、hsa-miR-372-3p、hsa-miR-495-3p、hsa-miR-520a-3p、hsa-miR-520b、hsa-miR-520d-3p、hsa-miR-520e、hsa-miR-613;调控辐射敏感组的miRNAs共5个:hsa-let-7c-5p、hsa-miR-137、hsa-miR-203a-3p、hsa-miR-34c-5p、hsa-miR-98-5p。结论利用生物信息学方法初步筛选到与非小细胞肺癌放疗敏感性相关的miRNAs。     

凋亡相关基因Bcl-XL、Bax表达与人卵巢癌细胞辐照凋亡效应的相关性研究

目的 :探讨 60Coγ射线作用下人卵巢癌细胞株A2 780及其顺铂耐药株A2 780 -cp凋亡相关基因Bcl -XL、Bax的表达及在辐照凋亡效应中作用。方法 :分别运用流式细胞技术 (FCM )及RT -PCR方法 ,分析A2 780、A2 780 -cp中Bcl -XL、Bax转录的相对表达与辐照诱导的细胞凋亡率的关系。结果 :A2 780 -cp细胞较其亲本株A2 780细胞有较高水平的Bcl-XL表达 ;辐照后 2h两者呈现不同程度的Bcl-XL、Bax的转录增加及Bcl-XL/Bax比率变化。结论 :不同p5 3基因状态卵巢癌细胞对辐照诱导的凋亡敏感性与肿瘤细胞的Bcl-XL、Bax表达及二者的比率有关     

Endoplasmic reticulum stress sensitizes human esophageal cancer cell to radiation

AIM:To investigate the role of endoplasmic reticulum(ER) stress in cancer radiotherapy and its molecular mechanism.METHODS:Tunicamycin(TM) was applied to induce ER stress in human esophageal cancer cell line EC109,and the radiosensitization effects were detected by acute cell death and clonogenic survival assay.Cell cycle arrest induced by TM was determined by flow cytometric analysis after the cellular DNA content was labeled with propidium iodide.Apoptosis of EC109 cells induced by TM was detected by annexin V staining and Western blotting of caspase-3 and its substrate poly ADP-ribose polymerase.Autophagic response was determined by acridine orange(AO) staining and Western blotting of microtubule-associated protein-1 light chain-3(LC3) and autophagy related gene 5(ATG5).In order to test the biological function of autophagy,specific inhibitor or Beclin-1 knockdown was used to inhibit autophagy,and its effect on cell apoptosis was thus detected.Additionally,involvement of the phosphatidylinositol-3 kinase(PI3K)/Akt/mammalian target of the rapamycin(mTOR) pathway was also detected by Western blotting.Finally,male nude mice inoculated subcutaneously with EC109 cells were used to confirm cell model observations.RESULTS:Our results showed that TM treatment enhanced cell death and reduced the colony survival fraction induced by ionizing radiation(IR),which suggested an obvious radiosensitization effect of TM.Moreover,TM and IR combination treatment led to a significant increase of G2/M phase and apoptotic cells,compared with IR alone.We also observed an increase of AO positive cells,and the protein level of LC3-II and ATG5 was induced by TM treatment,which suggested an autophagic response in EC109 cells.However,inhibition of autophagy by using a chemical inhibitor or Beclin-1 silencing led to increased cell apoptosis and decreased cell viability,which suggested a cytoprotective role of autophagy in stressed EC109 cells.Furthermore,TM treatment also activated mTORC1,and in turn reduced Akt phosphorylation,which sugge     

Chemosensitivity and radiosensitivity profiles of four new human epithelial ovarian cancer cell lines exhibiting genetic alterations in BRCA2, TGFbeta-RII, KRAS2, TP53 and/or CDNK2A

To address the cellular basis for the response to ovarian cancer treatment, we characterized the chemosensitivity and radiosensitivity of four human epithelial ovarian cancer cell lines that harbor different genetic alterations. The TOV-21G, TOV-81D, OV-90, and TOV-112D cell lines were derived from ovarian tumors (TOV) or ascites (OV) from chemotherapy- and radiotherapy-naive patients and were characterized by their mutation spectrum of BRCA2, TGF-RII, KRAS2, TP53, and CDKN2A. Cells were monitored for survival following exposure at various concentrations to different cytotoxic agents including cisplatin, camptothecin or paclitaxel or to different doses of -irradiation. At the lowest doses, the TGF-RII-mutated and KRAS2-mutated cell line, TOV-21G, and the BRCA2-mutated cell line, TOV-81D, demonstrated a significantly higher sensitivity to cisplatin and -irradiation than the TP53-mutated cell lines, TOV-112D and OV-90. At higher doses, differences between the TP53-mutated lines were observed with TOV-112D being less sensitive to cisplatin than OV-90 that also harbors a CDNK2A mutation. All cell lines were similarly sensitive to high doses of -irradiation. In contrast, sensitivity to camptothecin or paclitaxel was not significantly different between all cell lines, irrespective of the mutation status of BRCA1, BRCA2, TGF-RII, KRAS2, TP53, and CDKN2A. The observed responses to treatment are consistent with the current knowledge concerning BRCA2, TGF-RII, KRAS2, TP53, and/or CDKN2A aberrant function.The first and second authors contributed equally to this work.     

"彗星"分析法检测人癌裸鼠移植瘤的放射敏感性

目的　探讨“彗星”分析法应用于人实体肿瘤放射敏感性检测的可能性。方法　应用“彗星”分析法 ,以尾力距之比 (RTM)作为终指标 ,检测人肺腺癌、人食管鳞癌和人鼻咽鳞癌等 3种人癌裸小鼠移植瘤的放射敏感性。裸小鼠移植瘤组织被消化并稀释成细胞浓度为 4× 10 4ml的单细胞悬液后 ,分成对照组 (0Gy)和不同剂量照射组 ,照射组分别给予 2、5、10和 15Gy的 6MVX射线的冰上照射 ,照射后立即进行“彗星”分析。结果　 3种人癌裸小鼠移植瘤细胞未照射时 (0Gy)的尾力矩 (TM)差异有显著性意义 (F =9.11,P <0 .0 1)。不同剂量 (2、5、10和 15Gy)照射后 ,3种人癌裸小鼠移植瘤细胞未做校正时的TM所反映放射敏感性由高到低的变化趋势依次为 :肺腺癌 >食管鳞癌 >鼻咽鳞癌 ,显然与临床一般印象不符 ;而经与对照组进行“本底”校正后的RTM所反映放射敏感性由高到低的变化趋势依次为 :食管鳞癌 >鼻咽鳞癌 >肺腺癌 ,则符合临床一般印象。结论　①应用“彗星”分析法检测实体肿瘤的放射敏感性 ,尾力矩必须进行“本底”校正 ,即扣除对照组本底误差后的尾力矩才能很好地反映实体肿瘤的放射敏感性差异。②“彗星”分析法可用于检测人体肿瘤组织的放射敏感性。