Enhancement of radiosensitivity of melanoma cells by pegylated gold nanoparticles under irradiation of megavoltage electrons

Purpose: Gold nanoparticles (GNP) have significant potential as radiosensitizer agents due to their distinctive properties. Several studies have shown that the surface modification of nanoparticles with methyl polyethylene glycol (mPEG) can increase their biocompatibility. However, the present study investigated the radiosensitization effects of mPEG-coated GNP (mPEG-GNP) in B16F10 murine melanoma cells under irradiation of 6?MeV Electron beam.

Materials and methods: The synthesized GNP were characterized by UV-Visible spectroscopy, dynamic light scattering, transmission electron microscopy, and zeta potential. Enhancement of radiosensitization was evaluated by the clonogenic assay at different radiation doses of megavoltage electron beams.

Results: It was observed that mPEG-GNP with a hydrodynamic size of approximately 50?nm are almost spherical and cellular uptake occurred at all concentrations. Both proliferation efficiency and survival fraction decreased with increasing mPEG-GNP concentration. Furthermore, significant GNP sensitization occurred with a maximum dose enhancement factor of 1.22 at a concentration of 30 μM.

Conclusions: Pegylated-GNP are taken up by B16F10 cancer cells and cause radiosensitization in the presence of 6?MeV electrons. The radiosensitization effects of GNP may probably be due to biological processes. Therefore, the underlying biological mechanisms beyond the physical dose enhancement need to be further clarified.     

miR-885-3p靶向AKT1对结直肠癌细胞HT-29放射敏感性的影响

目的 探究miR-885-3p对结直肠癌细胞HT-29放射敏感性的影响以及作用机制。方法 荧光定量PCR检测经不同剂量（0、2、4、6、8 Gy）X射线照射后HT-29细胞中miR-885-3p的表达量;建立过表达miR-885-3p细胞株,功能试验探讨其对HT-29细胞放射敏感性的影响;生物信息学预测miR-885-3p下游调控的靶基因,双荧光素酶报告基因法进一步验证;上调和下调miR-885-3p表达量探讨miR-885-3p与靶基因丝苏氨酸蛋白激酶1（AKT1）表达量的调控关系;慢病毒转染敲减AKT1表达量,观察其对HT-29细胞放射敏感性的影响;共转染miR-885-3p模拟物,探讨过表达AKT1对miR-885-3p诱导的HT-29细胞放射敏感性的影响。结果 miR-885-3p在放射诱导的HT-29细胞中表达上调（F=46.64,P<0.05）;过表达miR-885-3p和敲减AKT1可通过抑制HT-29细胞存活、促进其凋亡,从而增强HT-29细胞放射敏感性（t=12.33、12.95,P<0.05）,放射增敏比（SER）分别为1.602和1.946;抑制miR-885-3p可通过促进HT-29细胞存活、抑制其凋亡从而促进HT-29细胞放射抵抗（t=11.94,P<0.05）,SER为0.839;AKT1是miR-885-3p下游靶基因;过表达AKT1反转miR-885-3p增强HT-29放射敏感性的作用,SER为0.680。结论 miR-885-3p通过直接靶向AKT1增加结直肠癌HT-29细胞放射敏感性,为提高临床结直肠癌放疗敏感性提供一个靶点。     

LncRNA CRNDE靶向miR-384影响结直肠癌细胞放射敏感性的研究

目的 研究长链非编码RNA （Lnc RNA）结直肠肿瘤差异表达基因（CRNDE）对结直肠癌细胞放射敏感性的影响及其机制。方法 以结直肠癌HT-29细胞作为体外研究对象,转染CRNDE shRNA,实时定量PCR测定干扰效果。以8 Gy X射线照射转染CRNDE shRNA后的HT-29细胞,四甲基偶氮唑盐（MTT）比色法和流式细胞术分别检测细胞增殖和凋亡水平。平板克隆实验检测放射敏感性。生物信息学软件预测CRNDE与miR-384有互补结合位点,荧光素酶报告系统鉴定靶向关系。将CRNDE shRNA和miR-384 inhibitor共转染至HT-29细胞中,以8 Gy剂量照射处理,MTT和流式细胞术检测细胞增殖和凋亡变化。结果 CRNDE shRNA能够降低HT-29细胞中CRNDE表达水平（1.00±0.08 vs. 0.42±0.06,t=10.051,P<0.05）。CRNDE shRNA和放射均可以抑制HT-29细胞增殖并诱导细胞凋亡,并且二者联合具有协同作用[凋亡率：（2.27±0.13）%、（23.58±2.35）%、（26.91±2.81）%、（36.84±3.24）%,F=24.66,P<0.05;吸光度（A）值：0.45±0.06、0.30±0.02、0.28±0.03、0.20±0.02,F=106.21,P<0.05]。CRNDE shRNA转染后可以提高HT-29细胞放射敏感性,放射增敏比为1.374。CRNDE靶向负调控miR-384表达。miR-384 inhibitor能够拮抗CRNDE shRNA对放射处理的结直肠癌细胞增殖抑制和凋亡促进的作用。结论 下调LncRNA CRNDE表达可增强结直肠癌细胞的放射敏感性,其作用机制与靶向负调控miR-384表达有关。     

PARP inhibitor attenuated colony formation can be restored by MAP kinase inhibitors in different irradiated cancer cell lines

Abstract

Purpose: Sensitizing cancer cells to irradiation is a major challenge in clinical oncology. We aimed to define the signal transduction pathways involved in poly(ADP-ribose) polymerase (PARP) inhibitor-induced radiosensitization in various mammalian cancer lines.

Materials and methods: Clonogenic survival assays and Western blot examinations were performed following telecobalt irradiation of cancer cells in the presence or absence of various combinations of PARP- and selective mitogen-activated protein kinase (MAPK) inhibitors.

Results: HO3089 resulted in significant cytotoxicity when combined with irradiation. In human U251 glioblastoma and A549 lung cancer cell lines, Erk1/2 and JNK/SAPK were found to mediate this effect of HO3089 since inhibitors of these kinases ameliorated it. In murine 4T1 breast cancer cell line, p38 MAPK rather than Erk1/2 or JNK/SAPK was identified as the main mediator of HO3089's radiosensitizing effect. Besides the aforementioned changes in kinase signaling, we detected increased p53, unchanged Bax and decreased Bcl-2 expression in the A549 cell line.

Conclusions: HO3089 sensitizes cancer cells to photon irradiation via proapoptotic processes where p53 plays a crucial role. Activation of MAPK pathways is regarded the consequence of irradiation-induced DNA damage, thus their inhibition can counteract the radiosenzitizing effect of the PARP inhibitor.     

Radiosensitization of ultrasmall GNP–PEG–cRGDfK in ALTS1C1 exposed to therapeutic protons and kilovoltage and megavoltage photons

Purpose: One of the promising radiosensitizers is the ultrasmall gold nanoparticle (GNP) with a hydrodynamic diameter?<3?nm. We studied functionalized ultrasmall GNPs (1.8?nm diameter) coated by polyethylene glycol (PEG) and conjugated with cyclic RGDfK (2.6?nm hydrodynamic diameter) for targeting of alpha(v) beta(3) integrin (α_vβ₃) in the murine ALTS1C1 glioma cell line.

Materials and methods: We investigated the uptake, toxicity and radiosensitivity of GNP–PEG–cRGDfKs in ALTS1C1 cells exposed to protons, kilovoltage photons and megavoltage photons. The in vitro uptake and toxicity of GNPs in the hepatocytes and Kupffer cells were assessed for murine AML12 hepatocyte and RAW 264.7 macrophage cell lines. The in vivo biodistribution of GNPs in the ALTS1C1 tumor model was tested using the inductively coupled plasma mass spectrometry.

Results: Results indicated GNPs accumulated in the cytoplasm with negligible toxicity for a moderate concentration of GNPs. Observed sensitizer enhancement ratios and dose enhancement factors are 1.21–1.66 and 1.14–1.33, respectively, for all radiations.

Conclusion: Ultrasmall GNP–PEG–cRGD can be considered as a radiosensitizer. For radiotherapy applications, the delivery method should be developed to increase the GNP uptake in the tumor and decrease the uptakes in undesirable organs.     

Radiation dose rate affects the radiosensitization of MCF-7 and HeLa cell lines to X-rays induced by dextran-coated iron oxide nanoparticles

Background and purpose: The aim of radiotherapy is to deliver lethal damage to cancerous tissue while preserving adjacent normal tissues. Radiation absorbed dose of the tumoral cells can increase when high atomic nanoparticles are present in them during irradiation. Also, the dose rate is an important aspect in radiation effects that determines the biological results of a given dose. This in vitro study investigated the dose-rate effect on the induced radiosensitivity by dextran-coated iron oxide in cancer cells.

Materials and methods: HeLa and MCF-7 cells were cultured in vitro and incubated with different concentrations of dextran-coated iron oxide nanoparticles. They were then irradiated with 6?MV photons at dose rates of 43, 185 and 370 cGy/min. The MTT test was used to obtain the cells’ survival after 48?h of irradiations.

Results: Incubating the cells with the nanoparticles at concentrations of 10, 40 and 80?μg/ml showed no significant cytotoxicity effect. Dextran-coated iron oxide nanoparticles showed more radiosensitivity effect by increasing the dose rate and nanoparticles concentration. Radiosensitization enhancement factors of MCF-7 and HeLa cells at a dose-rate of 370 cGy/min and nanoparticles’ concentration of 80?μg/ml were 1.21?±?0.06 and 1.19?±?0.04, respectively.

Conclusion: Increasing the dose rate of 6?MV photons irradiation in MCF-7 and HeLa cells increases the radiosensitization induced by the dextran-coated iron nanoparticles in these cells.     

Radiation-induced DNA damage response and resistance in colorectal cancer stem-like cells

Abstract

Purpose: Radiation therapy is an integral part of current treatment modality for colorectal cancer. Recent studies have revealed the presence of cancer stem-like cells (CSCs) population, in different tumors are responsible for therapeutic resistance and disease relapse, including colorectal cancer with poorer survival rate. Hence, characterization of the effect of Ionizing Radiation (IR) in colorectal cancer may serve to explain possible mechanisms.

Material and methods: Parental HCT116 and HCT-15 cells and derived colonospheres were irradiated and dose was optimized based on cell survival assay and cell cycle analysis. DNA damage response (DDR) was elucidated by γH2AX foci formation, COMET assay, and ATM, p-ATM, ERCC1 expression post-treatment. The expression level of developmental marker (β-catenin), CSC markers (CD44, KLF4) and telomeric components (TRF2, RAP1, hTERT) were evaluated.

Results: We observed cell survival was more in colonospheres post-irradiation and also exhibited decreased γH2AX foci, olive tail moment, increased ERCC1, and p-ATM expression than its parental counterpart which corresponds to efficient DDR. Differential expression of developmental marker, CSC markers, and telomeric components were observed after irradiation.

Conclusion: This study highlighted the presence of CSC phenotype in colonospheres having increased DNA repair capacity. Differential expression of developmental marker, CSC markers and telomeric components between parental and colonospheres may contribute in radio-resistance property of CSCs.     

Increased radiotoxicity in two cancerous cell lines irradiated by low and high energy photons in the presence of thio-glucose bound gold nanoparticles

Purpose: Gold nanoparticles modified by thio-glucose are believed to increase the toxicity of radiotherapy in human malignant cells. We report the effect of thio-glucose bound gold nanoparticles (Glu-G nanoparticles), 16?nm in size, on two human lung (QU-DB) and breast (MCF7) cancer cell lines combined with kilo and megavoltage X-rays.

Materials and methods: The shape and surface characteristics, the size distribution and light absorption spectrum of the prepared nanoparticles were measured by transmission electron microscopy, dynamic light scattering, and ultraviolet-visible spectrophotometry, respectively. The cell uptake was assayed using the atomic absorption spectrometry. Mitochondrial activity, colony formation, and comet assays were applied to assess and compare the enhanced radiotoxicity of 100?KV and 6?MV X-rays, when combined with Glu-G nanoparticles.

Results: Glu-G nanoparticles had no significant toxicity for MCF7 and QU-DB cells up to 100 micromolar concentration. Compared to radiation alone, the intracellular uptake of Glu-G nanoparticles resulted in increased inhibition of cell proliferation by 64.1% and 38.7% for MCF7 cells, and 64.4% and 32.4% for QU-DB cells by 100 kVp and 6?MV X-rays, respectively. Comet assay confirmed an increase of DNA damage as a result of combination of 6?MV photons with Glu-G nanoparticles.

Conclusion: Glu-G nanoparticles have remarkable potential for enhancing radiotoxicity of both low and high energy photons in MCF7 and QU-DB cells.     

抑制FOXD1基因表达增强结直肠癌细胞HCT116的放射敏感性

目的 研究抑制FOXD1基因的表达对结直肠癌细胞放射敏感性的影响。方法 采用实时荧光定量聚合酶链反应（qRT-PCR）和Western blot检测人结直肠癌组织和细胞中FOXD1 mRNA和蛋白的表达。对结直肠癌HCT116细胞行梯度剂量（0、2、4、6 Gy）X射线照射,qRT-PCR和Western blot检测各组细胞中FOXD1的表达。将siRNA阴性对照和FOXD1 siRNA转染至结直肠癌细胞中,分别记为si-NC组和si-FOXD1组,经4 Gy的X射线照射处理后记为si-NC+4 Gy组和si-FOXD1+4 Gy组,Western blot检测各组细胞中FOXD1的表达,四甲基偶氮唑盐（MTT）法检测各组细胞的增殖活性,克隆形成实验检测各组细胞存活率,采用TECT DNA-PK试剂盒检测各组细胞中DNA-PK活性,将转染的结直肠癌细胞接种于BALB/c裸鼠建立移植瘤模型,进行射线照射后,检测各组肿瘤的体积和质量变化。结果 与癌旁正常组织相比,结直肠癌组织中FOXD1 mRNA和蛋白的表达均显著增加（t=5.579、4.816,P<0.05）,与结肠黏膜上皮细胞NCM460相比,结直肠癌细胞株中FOXD1 mRNA（t=5.85~17.62,P<0.05）和蛋白（t=9.04~11.42,P<0.05）表达均显著升高。结直肠癌细胞HCT116中FOXD1的表达量随着放射剂量增加而升高,呈现剂量依赖性,差异有统计学意义（t=9.13~44.15,P<0.05）。转染si-FOXD1能够有效地抑制结直肠癌细胞中FOXD1的表达（t=10.51,P<0.05）,FOXD1敲低后能够抑制结直肠癌细胞的增殖活性（t=10.41,P<0.05）,提高结直肠癌细胞的放射敏感性,放射增敏比为1.797,降低放射诱导的DNA-PK的活性（t=6.20,P<0.05）。抑制FOXD1的表达经射线照射后,裸鼠种植瘤体积和重量明显减小（t=11.29、3.69,P<0.05）。结论 抑制FOXD1基因的表达能够提高结直肠癌细胞的放射敏感性,抑制结直肠癌裸鼠移植瘤的生长,可为改善放射治疗对结直肠癌患者的治疗效果提供潜在的靶向基因。     

MLN4924 and 2DG combined treatment enhances the efficiency of radiotherapy in breast cancer cells

Purpose: Two-deoxy-D-glucose (2DG) causes cytotoxicity in the cancer cells by disrupting the thiol metabolism, and MLN4924 inactivates the SCF E3 ligase and so causes the accumulation of its substrates which trigger apoptosis and hence might enhance the efficiency of radiotherapy and overcame on the radioresistance of the cancer cells.

Materials and methods: SKBR3 and MCF-7 breast cancer cells were treated with 500?μM 2DG and/or MLN4924 (30, 100, 200 and 300?nM), and in combination in the presence and absence of 1, 1.5 and 2?Gy gamma irradiation. The effects of the treatments – 2DG, MLN4924, irradiation alone and combined – on MCF-7 and SKBR3 cell lines were evaluated by MTT assay, TUNEL assay, cell death detection, Q-PCR for caspase-3 and Bcl-2 expression analysis, and finally clonogenic survival assay.

Results: The treatments enhanced the further radio cytotoxicity via inducing the apoptosis cell signaling gene, caspase-3. The 2DG and MLN4924 treatments could act as a radiosensitizer, especially on the SKBR3 cells, and further sensitized the cells with a sensitivity enhancement ratio (SER) of 1.41 and 1.27 in SKBR3 and MCF-7 cells, respectively.

Conclusion: The combined chemo-radiotherapy might improve the breast cancer treatment outcome.     

Biological outcomes of γ-radiation induced DNA damages in breast and lung cancer cells pretreated with free radical scavengers

Abstract

Purpose: Investigation of effects on DNA of γ-irradiated human cancer cells pretreated with free radical scavengers is aimed to create reference data which would enable assessment of the relative efficiency of high linear energy transfer (LET) radiations used in hadron therapy, i.e. protons and carbon ions.

Materials and methods: MCF-7 breast and HTB177 lung cancer cells are irradiated with γ-rays. To minimize indirect effects of irradiation, dimethyl sulfoxide (DMSO) or glycerol are applied as free radical scavengers. Biological response to irradiation is evaluated through clonogenic cell survival, immunocytochemical and cell cycle analysis, as well as expression of proteins involved in DNA damage response.

Results: Examined cell lines reveal similar level of radioresistance. Application of scavengers leads to the rise of cell survival and decreases the number of DNA double strand breaks in irradiated cells. Differences in cell cycle and protein expression between the two cell lines are probably caused by different DNA damage repair mechanisms that are activated.

Conclusion: The obtained results show that DMSO and glycerol have good scavenging capacity, and may be used to minimize DNA damage induced by free radicals. Therefore, they will be used as the reference for comparison with high LET irradiations, as well as good experimental data suitable for validation of numerical simulations.     

1-Methylxanthine enhances the radiosensitivity of tumor cells

Purpose:?To determine the efficacy of a caffeine derivative 1-methylxanthine (1-MTX) in increasing radiosensitivity of cancer cells and elucidate the underlying mechanisms in vitro.

Materials and methods:?RKO human colorectal cancer cells carrying wild type protein 53 kDa (p53) were incubated with 3 mM 1-MTX for 30 min, exposed to 4 Gy ionizing radiation, and further incubated with 1-MTX for three days. The clonogenic cell death was determined, and the cell cycle distribution and apoptosis were studied with flow cytometry at different times after irradiation. The DNA double strand break (DNA DSB) was examined using phosphorylated Histone2A (γ-H2AX) foci formation, and the expression/activity of checkpoint 2 kinase (Chk2), cell division cycle 25 (Cdc25) phosphatase and cyclin B1/Cdc2 kinase were also investigated using western blotting and in vitro kinase assays.

Results:?The treatment with 3 mM 1-MTX increased the radiation-induced clonogenic and apoptotic cell death. The radiation-induced phosphorylation of Chk2 and Cdc25c and the radiation-induced increase in the cyclin B1/Cdc2 kinas activity were little affected by 1-MTX. The radiation-induced G2/M arrest was only slightly shortened and the expression of radiation-induced γ-H2AX was markedly prolonged by 1-MTX.

Conclusions:?1-MTX significantly increased the radiosensitivity of RKO human colorectal cancer cells carrying wild type p53 mainly by inhibiting the repair of radiation-induced DNA DSB without causing significant alteration in radiation-induced G2/M arrest. Such a radiosensitization occurred at 1-MTX concentrations almost non-toxic to the target tumor cells.     

Radiosensitization by Cisplatin of RIF1 Tumour Cells in Vitro

Summary

the ability of cis-diamminedichloroplatinum (II) (c-DDP) to enhance radiation-induced cell killing was tested on oxic RIF1 tumour cells in monolayer culture. Marked radiosensitization of the survivors of a 1 h drug treatment was found with all c-DDP doses tested, with enhancement ratios increasing from 1·2 to 2·2 with increasing drug dose. Isobologram analyses showed that the interactions of c-DDP with X-rays were supra-additive.

To test whether part of the enhancement was due to a selection of subpopulations, the diploid and tetraploid RIF1 cells, which normally coexist in culture, were separated by (a) unit gravity velocity sedimentation, and (b) by developing diploid and tetraploid clones. Both methods showed that there was little differnce in either drug sensitivity or radiation sensitivity between diploid and tetraploid cells. DNA histograms obtained by flow cytometry showed little or no cycle progression during the 1 h drug treatment. These data indicate that the radiosensitization was not the result of the drug exposure leaving cells in a radiosensitive phase. The observed radiosensitization, therefore, appears to have resulted from a true drug/X-ray interaction.     

Biological characteristics of gene expression features in pancreatic cancer cells induced by proton and X-ray irradiation

Abstract

Background: Radiation therapy is an important alternative treatment for advanced cancer. The aim of the current study was to disclose distinct alterations of the biological characteristics of gene expression features in pancreatic cancer cells, MIAPaCa-2, following proton and X-ray irradiation.

Materials and methods: Using cDNA microarray, we examined the gene expression alterations of MIAPaCa-2 cells following proton or X-ray irradiation. We also isolated the surviving MIAPaCa-2 cells after irradiation and analyzed their gene expression profiles.

Results: Although the cytocidal effects of both types of irradiation were similar at sufficient doses in vitro and in vivo, the affected gene expression profile alterations of MIAPaCa-2 cells irradiated with protons were distinct from those irradiated with X-ray. Interestingly, clustering analysis of gene expression of the surviving MIAPaCa-2 cells was also completely discernible between the two types of irradiation. However, a similar cytocidal effect was still observed in the proton- and X-ray-irradiated surviving cells after re-irradiation, commonly showing biological effects related to apoptosis and cell cycle processes.

Conclusions: Proton irradiation treatment for pancreatic cancer provides the distinct biological effect of steady gene expression alterations compared to X-ray irradiation; however, surviving cells from both types of irradiation were still susceptible to the cytocidal effects induced by proton re-irradiation treatment.     

Low-dose non-targeted radiation effects in human esophageal adenocarcinoma cell lines

Purpose: To investigate non-targeted radiation effects in esophageal adenocarcinoma cell lines (OE19 and OE33) using human keratinocyte and colorectal cancer cell reporters following γ-ray exposure.

Materials and methods: Both clonogenic assays and ratiometric calcium endpoints were used to check for the occurrence of bystander signals in reporter cells.

Results: We report data suggesting that γ-irradiation increases cell killing over the expected linear quadratic (LQ) model levels in the OE19 cell line exposed to doses below 1?Gy, i.e. which may be suggestive to be a low hyper-radiosensitive (HRS) response to direct irradiation. Both EAC cell lines (OE19 and OE33) have the ability to produce bystander signals when irradiated cell conditioned medium (ICCM) is placed onto human keratinocyte reporters, but do not seem to be capable of responding to bystander signals when placed on their autologous reporters. Further work with human keratinocyte reporter models showed statistically significant intracellular calcium fluxes following exposure of the reporters to ICCM harvested from both EAC cell lines exposed to 0.5?Gy.

Conclusion: These experiments suggest that the OE19 and OE33 cell lines produce bystander signals in human keratinocyte reporter cells. However, the radiosensitivity of the EAC cell lines used in this study cannot be enhanced by the bystander response since both cell lines could not respond to bystander signals.     

西妥昔单抗对人舌癌细胞系Tca8113的放射增敏作用

目的 探讨西妥昔单抗( C225)对人舌癌Tca8113细胞系的放射增敏作用.方法 体外培养人舌癌Tca8113细胞系,C225组用西妥昔单抗(C225) 100 nmol/L处理后,6MVX射线不同剂量(0、2、4、6、8和10 Gy)照射C225组和单纯照射组,照射后用细胞计数法及四甲基偶氮唑盐比色法(MTT)检测细胞增殖率,集落形成实验计算克隆数,采用流式细胞仪检测细胞周期、细胞凋亡.结果 不同照射剂量时C225+照射组的细胞生长抑制率均高于单纯照射组(t=-15.6～-3.0,P ＜0.05);C225+照射组的放射生物学参数值(D0、Dq、N、SF2)较单纯照射组低.C225+照射组的SER为1.353;C225+照射组的G0/G1期细胞比例在4、6、8 Gy时均高于单纯照射组(t=-7.64、-7.89、-4.78,P＜0.05).随着照射剂量升高,C225+照射组和单纯照射组的细胞早期凋亡率皆先升高后下降,4 Gy时两组差异最大[(7.96±0.36)％和(4.13±0.29)％,t- 12.75,P＜0.01].结论 C225对人舌癌Tca8113细胞系具有放射增敏作用.C225可能通过Go/G.期阻滞和诱导凋亡来增加人舌癌Tca8113细胞系的放射敏感性.     

The role of iron oxide nanoparticles in the radiosensitization of human prostate carcinoma cell line DU145 at megavoltage radiation energies

Abstract

Purpose: To examine the radiosensitizing effects of iron oxide nanoparticles in the presence of 6 MV (megavoltage) X-ray radiation.

Materials and methods: Iron oxide nanoparticles with two different modifications – dextran coating (plain) and amino-group dextran coating – were used. The rate of iron oxide penetration was monitored using Prussian blue staining, magnetic resonance imaging and atomic adsorption spectroscopy. The effect of iron oxide on the viability of cells was determined using trypan blue dye exclusion assay followed by evaluating the cytotoxicity effect of amino-group iron oxide nanoparticles and ionizing radiation. Radiation dose enhancement studies were carried out on DU145 human prostate carcinoma cell line with 1 mg/ml amino-group iron oxide nanoparticles and different doses of 6 MV X-ray radiation.

Results: The uptake of amino-group coated nanoparticles by DU145 cells was significantly more than the plain nanoparticles. In addition, cell viability was decreased with the increase of iron oxide concentration. The dose enhancement factor (DEF) is approximately 1.2 at different doses in the range of 2–8 Gy of 6 MV X-ray radiation.

Conclusions: It was demonstrated that iron oxide nanoparticles with the appropriate surface modifications can enter the DU145 cells and it can be used as a cell sensitizer to megavoltage ionizing radiations in radiation therapy.     

Photodynamic action of palmatine hydrochloride on colon adenocarcinoma HT-29 cells

Palmatine hydrochloride (PaH) is a natural active compound from a traditional Chinese medicine (TCM). The present study aims to evaluate the effect of PaH as a new photosensitizer on colon adenocarcinoma HT-29 cells upon light irradiation. Firstly, the absorption and fluorescence spectra of PaH were measured using a UV–vis spectrophotometer and RF-1500PC spectrophotometer, respectively. Singlet oxygen (¹O₂) production of PaH was determined using 1, 3-diphenylisobenzofuran (DPBF). Dark toxicity of PaH was estimated using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cellular uptake of PaH in HT-29 cells was detected at different time intervals. Subellular localization of PaH in HT-29 cells was observed using confocal laser fluorescence microscopy. For photodynamic treatment, HT-29 cells were incubated with PaH and then irradiated by visible light (470 nm) from a LED light source. Photocytotoxicity was investigated 24 h after photodynamic treatment using MTT assay. Cell apoptosis was observed 18 h after photodynamic treatment using a flow cytometry with Annexin V/PI staining. Results showed that PaH has an absorption peak in the visible region from 400 nm to 500 nm and a fluorescence emission peak at 406 nm with an excitation wavelength of 365 nm. PaH was activated by the 470 nm visible light from a LED light source to produce ¹O₂. Dark toxicity showed that PaH alone treatment had no cytotoxicity to HT-29 cancer cells and NIH-3T3 normal cells after incubation for 24 h. After incubation for 40 min, the cellular uptake of PaH reached to the maximum and PaH was located in mitochondria. Photodynamic treatment of PaH demonstrated a significant photocytotoxicity on HT-29 cells. The rate of cell death increased significantly in a PaH concentration-dependent and light dose-dependent manner. Further evaluation revealed that the early and late apoptotic rate of HT-29 cells increased remarkably up to 21.54% and 5.39% after photodynamic treatment of PaH at the concentration of 5 μM and energy density of 10.8 J/cm². Our findings demonstrated that PaH as a naturally occurring photosensitizer has potential in photodynamic therapy on colon adenocarcinoma.     

Caffeine-increased radiosensitivity is not dependent on a loss of G2/M arrest or apoptosis in bladder cancer cell lines.

PURPOSE: Bladder cancer cell lines UCRU-BL-13, UCRU-BL-17/2 and UCRU-BL-28, with differing p53 status and molecular responses to irradiation, were used to investigate possible mechanisms for caffeine-induced radiosensitization. MATERIALS AND METHODS: After treatment with caffeine and exposure to X-radiation, radiosensitivity was determined by clonogenic assay. Cell-cycle arrest and apoptosis were measured by flow cytometry. RESULTS: Both BL-13 and BL-28 cells (each expressing p53 with a wild-type sequence) fail to arrest at the G2 checkpoint after radiation, but nevertheless caffeine did induce radiosensitization. In contrast, in BL-17/2 cells (expressing p53 with a point mutation in codon 280), caffeine treatment abrogated the radiation-induced G2 arrest but was not accompanied by radiosensitization. No effects on radiosensitivity were seen in RT112 cells (expressing a functionally defective p53) at low caffeine doses (2 mM), but at higher doses (4 mM and 10 mM) caffeine caused both abrogation of radiation-induced G2 arrest and radiosensitization. In none of the cell lines examined did caffeine treatment and/or irradiation result in apoptosis. CONCLUSIONS: In contrast with previous studies, the data suggest that radiosensitization induced by caffeine is not dependent on abrogation of G2 arrest or the induction of apoptosis, and is not selective for cells expressing p53 proteins with mutations.