miR-339-5p inhibits NUDT5 and enhances radiosensitivity of lung cancer A549 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the influence of miR-339-5p on the radio-sensitivity of lung cancer A549 cells by regulating the expression of Nudix hydrolase 5 (NUDT5). Methods: X-ray-resistant lung cancer A549 cells (RA549) were induced by treatment with low concentration gradient increment combined with large dose intermittent shock in vitro. The expression level of miR-339-5p in hu‐ man normal lung epithelial cells (BEAS-2B) and lung cancer cell lines (A549, L78, H1299, H460 and RA549 cells) was detected by qP‐ CR. According to the treatment, RA549 cells were divided into NC group, 5Gy group (treatment with 5Gy X-ray), 5Gy+miR-339-5p mimic group, 5Gy+si-NUDT5 group and 5Gy+si-NUDT5+miR-339-5p inhibitor group. CCK-8 assay, Annexin V-FITC/PI double stain‐ ing flow cytometry and WB were used to detect the proliferation, apoptosis and the protein expressions of NUDT5, γ-H2AX and H2AX in each group. The targeting relationship between mir-339-5p and NUDT5 was detected by Dual-luciferase reporter gene system. Results: The expression of miR-339-5p in lung cancer cell lines was significantly lower than that in BEAS-2B cells, with the lowest ex‐pression level in RA549 cells (all P<0.05). NUDT5 was the target gene of miR-339-5p. Compared with the NC group, the prolifera‐ tion activity and NUDT5 expression of RA549 cells in the 5 Gy group were significantly reduced (all P<0.01), and the apoptosis rate was significantly increased (P<0.01). Compared with the 5 Gy group, the proliferation activity of RA549 cells in the 5 Gy+miR-339-5p mimic group was significantly reduced (P<0.05), the apoptosis rate ([12.97±1.48]% vs [5.21±0.62]%, P<0.01) and the expression level of γ-H2AX (P<0.05) were significantly increased; the expression of NUDT5 (t=7.58, P<0.01) and cell proliferation activity (t=6.58, P<0.01) of RA549 cells in the 5 Gy+si-NUDT5 group were significantly reduced, while the apoptosis rate ([11.21±1.06]% vs [5.54±0.44%, P<0.01) and the expression of γ-H2AX (P<0.01) were significantly increased; and the above indicators in 5 Gy+si-NUDT5+miR-339-5p inhibitor group showed insignificant difference from the 5 Gy group. Conclusion: Overexpression of miR-339-5p enhances the radio-sensitivity of X-ray-resistant lung cancer A549 cells by targetedly down-regulating NUDT5 expression.

The knockdown of miR-23b enhances radio-sensitivity of lung cancer A549 cell via targeting PTEN / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of miR-23b/PTEN molecular axis on radio-sensitivity of lung cancerA549 cells and its mechanism. Methods: Lung cancer cell lines NCI-H1650, NCI-H175, Calu-1, LT-P-A-2, MSTO-211H, A549 and human normal lung epithelial cell line BEAS-2B were selected. The expression level of miR-23b in lung cancer cell lines was detected by qPCR. Dual-luciferase reporter gene assay was used to verify the relationship between miR-23b and PTEN. Plasmids miR-23b mimics, miR-23b inhibitor and pcDNA3.1-PTEN were transfected intoA549 cells by lipofection; PTEN expression levels in cells was detected by WB. CCK-8, Transwell andAnnexin V-FITC/PI staining flow cytometry were used to detect the effect of miR-23b/PTEN axis on proliferation, invasion and apoptosis ofA549 cells treated with 60Co γ-ray. Results: miR-23b was upregulated in lung cancer cell lines with the highest expression in A549 cells (P<0.05 or P<0.01). Knockdown of miR-23b reversed the inhibitory effect of 3 Gy 60Co γ-rays on proliferation and invasion of A549 cells, and induced apoptosis (P<0.05 or P<0.01). Dual-luciferase reporter gene assay results confirmed that miR23b could negatively regulate PTEN (P<0.05). Furthermore, knockdown of miR-23b up-regulated PTEN expression level, and furhter enhanced the inhibitory effect of 3 Gy 60Co γ-ray on the proliferation and invasion of A549 cells as well as induced apoptosis of A549 cells (P<0.05 or P<0.01). Conclusion: Knockdown of miR-23b can enhance the radio-sensitivity of A549 cells, the mechanism of which is that 60Co γ-ray down-regulates the inhibitory effect of miR-23b on PTEN, thereby inhibiting the proliferation, invasion and inducing apoptosis ofA549 cells.

miR-424 modulates radio-sensitivity of breast cancer cells via targeting HMGA1 / 中国肿瘤生物治疗杂志

@# Objective: To explore the effect of miR-424/HMGA1 (high mobility proteinA1) axis on the radio-sensitivity of breast cancer cells and the possible mechanism. Methods:Atotal of 50 cases of breast cancer tissues from patients, who underwent surgical resection at the Department of Oncological Radiotherapy, Wuxi Fourth People’s Hospital from April 2014 to April 2017, were collected for this study. Real-time quantitative polymerase chain reaction (qPCR) and Western blotting were performed to evaluate the mRNA and protein expressions of miR-424 and HMGA1 in breast cancer tissues of radiation sensitive and insensitive patients. After being treated with different doses of 60Co γ-ray radiation (0, 2, 4, 6 and 8 Gy), the expression changes of miR-424 and HMGA1 in breast cancer MDA-MB-468 cells were observed. Subsequently, miR-424 mimic/inhibitor and pcDNA-HMGA1 were transfected into MDA-MB-468 cells, and the effect of miR-424 on cell proliferation, invasion and apoptosis of radiation-treated MDA-MB-468 cells were evaluated by colony formation assay, MTT assay, Transwell assay and Annexin V-FITC/PI double staining flow cytometry assay, respectively. Furthermore, dual luciferase reporter gene assay was used to verify whether HMGA1 was a target gene of miR-424. Results: The patients in radio-sensitive group exhibited higher miR-424 expression but lower HMGA1 expression than the patients in insensitive group (all P<0.01). Compared with the cells treated with 0, 2 and 4 Gy radiation, the cells treated with 6 and 8Gy radiation exhibited significantly higher apoptosis rate and miR-424 expression but lower HMGA1 expression and cell invasion (all P<0.01). Moreover, luciferase reporter gene assay confirmed that miR-424 down-regulated HMGA1 expression. Mechanistically, miR-424 significantly inhibited cell proliferation, invasion and induced apoptosis of MDA-MB-468 cells (all P<0.01) via targeted down-regulating HMGA1, and further upregulated the radio-sensitivity of breast cancer cells. Conclusion: miR-424/HMGA1 axis regulates the radio-sensitivity of breast cancer, and over-expression of miR-424 may increase the sensitivity of MDA-MB-468 cells to γ-ray radiation therapy.

Licorice enhances radio-sensitivity of nasopharyngeal carcinoma CNE-2 cells by affecting autophagy / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of licorice on radio-sensitization of nasopharyngeal carcinoma CNE-2 cells and its mechanism. Methods: The radio-resistant nasopharyngeal carcinoma cell line (CNE-2-RR) was constructed and cultured in vitro. MTT assay was used to detect the effect of different concentrations of licorice on the proliferation activity of nasopharyngeal carcinoma cells. The changes of autophagosome in CNE-2-RR cells after licorice treatment were observed by transmission electron microscopy (TEM). Western blotting was used to detect the effect of licorice on the level of autophagy protein in CNE-2-RR cells. Single cell gel electrophoresis (comet assay) was used to detect the DNAdamage and repair of different groups of CNE-2-RR cells. Flow cytometry was used to detect the apoptosis rate of CNE-2-RR cell line. Results: Low-radiation resistant CNE-2-RR cell line was successfully constructed; MTT assay showed that 20 mmol/L licorice exhibited highest inhibition on CNE-2-RR cells (58.86 ± 5.02)%. Transmission electron microscopy showed increased autophagicbody and abnormal mitochondria and nuclei morphology in CNE-2-RR cells after treatment. Western blotting showed that autophagic protein LC3-II level was increased and LC3-I level was decreased in CNE-2-RR cells (P < 0.05). The results of single cell gel electrophoresis showed that the length of comet tail distance of CNE-2-RR cells after licorice treatment was higher than that of the control group (P<0.05), indicating weakened repair ability of DNA damage. Conclusion: Licorice enhances the radio-sensitivity of CNE-2-RR cells by influencing autophagy and DNArepair ability.

Influence of EMT in Radiosensitivity of PC-9:an EGFR- mutant NSCLC cell line / 实用医学杂志

Objective To explore the influence of epithelial mesenchymal transition(EMT)in radiosensi-tivity of EGFR-mutant NSCLC cell with acquired resistance to EGFR-TKI. Methods In this study,EGFR-mutant human lung adenocarcinoma cell line PC-9 and Gefitinib acquired resistance cell line PC-9/AB were used. Western blot was used to assess EMT. Wound healing migration assay was tested. CCK8,colony formation and flow cytome-try were used to evaluate survival fraction ,as the sensitivity to irradiation. Results PC-9/AB displayed radioresis-tance(P<0.05). When EMT was reversed with CDH1,its radiosensitivity was significantly higher than PC-9/AB (P < 0.05). PC-9/TGF also displayed radioresistance (P < 0.05),as well as EMT phenotype presented. Conclusion EMT enhanced the radioresistance of EGFR-mutant NSCLC cell with acquired resistance to EGFR-TKI, possibly through TGF-βpathway.

Effect of platycodin D on radiosensitivity of human hepatoma cell line and related mechanisms of action / 中华肝脏病杂志

Objective@#To investigate the effect of platycodin D on the radiosensitivity of human hepatoma cell lines HepG2 and SMMC-7721 and related mechanisms of action.@*Methods@#MTT assay was used to analyze the effect of different concentrations of platycodin D with different treatment times on cell viability. The cells were pretreated with 5 μg/ml platycodin D for 24 hours followed by X-ray irradiation at different radiation doses. Colony-forming assay was used to measure the radiosensitizing effect of platycodin D on cells. The quasi-threshold dose (Dq), mean lethal dose (Do), extrapolation number (N), sensitizer enhancement ratio (SER), and survival fraction (SF) at different radiation doses were calculated, and the multi-target single-hit model was used to fit the cell survival curve according to the formula SF = l-(l-e-D/D0)N. Flow cytometry was used to investigate the distribution of cell cycle, and Western blotting was used to measure the changes in the protein expression of phosphorylated phosphatidylinositol 3’-kinase (pPI3K), phosphorylated protein kinase (pAkt), nuclear factor-κB (NF-κB), and phosphorylated nuclear factor inhibiting protein (pIκBα). A one-way analysis of variance, the t-test, or the least significant difference test was used for statistical analysis based on the type of data.@*Results@#Platycodin D reduced the viability of HepG2 and SMMC-7721 cells in a dose-dependent manner; the IC50 value for HepG2 cells was 24.2 ± 0.61 μg/ml at 24 hours and 7.68 ± 0.46 μg/ml at 48 hours, and that for SMMC-7721 cells was 23.8 ± 0.57 μg/ml at 24 hours and 8.63 ± 0.86 μg/mL at 48 hours. After the combined treatment with platycodin D and irradiation, there were significant reductions in Dq (P = 0.002), Do (P = 0.002), and N value (P = 0.003), the survival curve markedly shifted to the left, and SER was 1.347 ± 0.04 in HepG2 cells and 1.418 ± 0.05 in SMMC-7721 cells. In addition, platycodin D significantly inhibited the increase in the proportion of cells in G2/M phase, the increases in the protein expression of pPI3k (P = 0.002), pAkt (P = 0.003), and NF-κB (P = 0.002), and the reduction in the protein expression of pIκBα (P = 0.003).@*Conclusion@#Platycodin D can increase the radiosensitivity of HepG2 or SMMC-7721 cells, possibly by enhancing the growth inhibition effect of irradiation and inhibiting the activation of the PI3k/Akt and NF-κB pathways.

Gold nanorods combined with internal radiotherapy in targeting therapy of tumor：recent progress in research / 介入放射学杂志

With the rapid development of nanotechnology, nano oncology medicine that is formed through the combination of nano science and oncology medicine has become an important field emerging in nano science. Because of its unique optical properties, the gold nanorod (GNR), that is a rod-shaped nano materials, has shown great potential applications in the biomedical field, especially, in GRNs thermotherapy, radio-sensitizing effect therapy and targeting therapy of the tumor. Being a newly-developed therapy, gold nanorods combined with internal radiotherapy has already obtained excellent durative effect in targeting therapy of tumor. This paper aims to make a comprehensive review about its mechanism and its clinical application and research situation in recent years.

Research on the progress of EGFR inhibitors in tumor inhibition and radio-sensitization / 中国肿瘤临床

Radiotherapy is important in cancer treatment, but improving the therapeutic effect of irradiation and decreasing its toxicity to normal human tissues is still a global problem. Epidermal growth factor receptor (EGFR) is a member of ErbB family and is an important transmembrane receptor with signal-transduction tyrosine kinase activity. EGFR can direct cellular migration, adhesion, proliferation, differentiation, and apoptosis, and plays a fundamental role in the development and growth of many types of human tumor cells. A series of preclinical studies showed that EGFR inhibitors can enhance the antitumor activity of ionizing radiation. EGFR inhibi-tors regulate radio-sensitization through multiple mechanisms, including cell cycle alterations, DNA repair modulation, and anti-angio-genesis. Reasonable application of EGFR inhibitors will effectively increase the radio-therapeutic effect, extend the local control of tu-mor, and improve a patient's quality of life.

Correlation of Nuclear Morphometry and AgNOR Score with Radiation Response in Squamous Cell Cancers of the Head and Neck: A Preliminary Study

Background: Prediction of radiation response before the completion of the radiotherapy schedule is challenging. Information about radiation response could help oncologist to choose the appropriate combination and sequence of therapies in the multidisciplinary management of cancer. Methods: The study involved 26 patients with squamous cell cancers of the head and neck region who received radiotherapy to a dose of 30 Gy in 10 fractions over a 2-week period as part of a split-course technique. Fine-needle aspiration cytology was performed on day 1 and day 5 of the schedule. The silver staining of the nuclear organiser region (AgNOR) and nuclear morphometric study were done on both days. Results: The median age of the patients was 44 years old. The primary tumours were distributed in the nasopharynx (n = 11), larynx and hypopharynx (n = 5), metastatic node (n = 4), and miscellaneous tumours were found in the head and neck sub sites (n = 6). The mean initial AgNOR score was 3.0, range 1.2–7.0. The median of nuclear and nucleolar diameters were 11.07 μm, range 7.70–16.6 μm, and 2.92 μm, range 1.09–11.66 μm, respectively. Patients with a pre-radiotherapy AgNOR score of greater than 2.5 were associated with disease progression and metastasis. However, the increased of nuclear diameter on day 5 compared with baseline predicted a good radiation response in patients (P = 0.016). Conclusion: Intra-radiotherapy nuclear morphometry combined with baseline AgNOR score could be a simple and useful tool for the prediction of radiation response in head and neck cancers.

TNF-alpha Downregulates E-cadherin and Sensitizes Response to gamma-irradiation in Caco-2 Cells / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The purpose of the present study was to assess the biological effects of TNF-alpha in Caco-2 well-differentiated colon adenocarcinoma cells and to determine radiation sensitivity in order to develop TNF-alpha into a cancer therapeutic agent. MATERIALS AND METHODS: A cell viability test was conducted via a colorimetric and colony forming assay after 1 day and 3 days of incubation with TNF-alpha. Western blotting analysis and immunofluorescence staining were conducted to explore TNF-alpha-induced morphological and molecular changes in the adhesion molecules, E-cadherin and claudin-4. The effects of gamma-irradiation at a dose of 2 Gy on cell survival were evaluated by a clonogenic assay. The molecular changes in apoptosis-regulatory proteins were assessed by Western blotting. RESULTS: Caco-2 cells were highly resistant to TNF alpha-induced cell death and 2 Gy of gamma-irradiation. However, we observed the downregulation of the adherens junctional protein, E-cadherin and translocation of tight junctional protein, claudin-4 from the membrane to the cytosol induced by TNF-alpha treatment which would indicate cell-cell junction disruptions. These alterations of junctional proteins influenced the regulation of cell death in response to 2 Gy of gamma-irradiation. The combined treatment of TNF-alpha with 2 Gy of gamma-irradiation reduced the survival of Caco-2 cells by down-regulating bcl-xl and activating JNK pathways. CONCLUSION: These results suggest that TNF-alpha might be potentially applied as a therapeutic agent in order to enhance sensitivity to 2 Gy of gamma-irradiation administered in radiotherapy for the treatment of human colon cancer.

Differential Expressions of Apoptosis-related Genes in Lung Cancer Cell Lines Determine the Responsiveness to Ionizing Radiation

Radiotherapy would be the choice of treatment for human cancers, because of high cost-effectiveness. However, a certain population of patients shows a resistance to radiotherapy and recurrence. In an effort to increase the efficacy of radiotherapy, many efforts were driven to find the genes causing the unresponsiveness to ionizing radiation. In this paper, we compared the gene expression profiles of two lung cancer cell lines, H460 and H1299, which showed differential responses to ionizing radiations. Each cell were irradiated at 2 Gy, and harvested after 0, 2, 4, 8, 12 and 24 hours to examine the expressions. Two-way ANOVA analysis on time-series experiments of two cells could select 2863 genes differentially expressed upon ionizing radiation among 32,321 genes in microarray (p<0.05). We classified these genes into 21 clusters by SOM clustering according to the interaction between cell types and time. Two SOM clusters were enriched with apoptosis-related genes in pathway analysis. One cluster contained higher levels of phosphatidyl inositol 3-phosphate kinase (PI3K) subunits in H1299, radio resistant cells than H460, radiosensitive cells. TRAIL receptors were expressed in H460 cells while the decoy receptor for TRAIL was expressed in H1299 cells. From these results, we could characterize the differential responsiveness to ionizing radiation according to their differential expressions of apoptosis-related genes, which might be the candidates to increase the power of radiotherapy.

Differential Expressions of Apoptosis-related Genes in Lung Cancer Cell Lines Determine the Responsiveness to Ionizing Radiation

Radiotherapy would be the choice of treatment for human cancers, because of high cost-effectiveness. However, a certain population of patients shows a resistance to radiotherapy and recurrence. In an effort to increase the efficacy of radiotherapy, many efforts were driven to find the genes causing the unresponsiveness to ionizing radiation. In this paper, we compared the gene expression profiles of two lung cancer cell lines, H460 and H1299, which showed differential responses to ionizing radiations. Each cell were irradiated at 2 Gy, and harvested after 0, 2, 4, 8, 12 and 24 hours to examine the expressions. Two-way ANOVA analysis on time-series experiments of two cells could select 2863 genes differentially expressed upon ionizing radiation among 32,321 genes in microarray (p<0.05). We classified these genes into 21 clusters by SOM clustering according to the interaction between cell types and time. Two SOM clusters were enriched with apoptosis-related genes in pathway analysis. One cluster contained higher levels of phosphatidyl inositol 3-phosphate kinase (PI3K) subunits in H1299, radio resistant cells than H460, radiosensitive cells. TRAIL receptors were expressed in H460 cells while the decoy receptor for TRAIL was expressed in H1299 cells. From these results, we could characterize the differential responsiveness to ionizing radiation according to their differential expressions of apoptosis-related genes, which might be the candidates to increase the power of radiotherapy.

Study on Anti-tumor Activities & Tumor Radio-sensitivity of 17a?-D-Homo-Ethynylestradiol-3-Acetate / 医学研究杂志

Objective The paper reports anti-tumor activities and tumor radio-sensitivity research of the novel Estrogen compound 17a?-D-Homo Ethynylestradiol-3-Acetate to U_ 14 and S_ 180 sarcomas;and overall test high-activity and low toxicity traits of 17a?-D-Homo Ethynylestradiol-3-Acetate by the indexes of thymus gland,spleen,etc.Methods The mouse cervical cancer U_ 14 was selected and implanted in IRM-2 mice for anti-tumor assay,and sarcoma S_ 180 was selected for tumor radio-sensitivity assay,the drug was administered to all mice by i.v.way.Results 17a?-D-Homo Ethynylestradiol-3-Acetate has obvious anti-tumor activity to U_ 14 tumor,the best inhibitory rates is U_ 14 64.3%,and17a?-D-Homo Ethynylestradiol-3-Acetate has hardly any influence to hematogenous system(spleen index),immune system(thymus index);Nevertheless,the positive control drug CCP has obvious damage to spleen index and thymus index(P