Glioblastoma (GBM) radiotherapy is hampered by intrinsic radioresistance. Current radiosensitizers face two unresolved hurdles: inability to dynamically traverse sequential physiological barriers of GBM and lack of multitargeted action against the pathways driving radioresistance. Here, we developed h-Pep-MTZ, a biobarrier-adaptive peptide-radiosensitizer addressing both. This system undergoes smart multistage transformations to overcome key delivery barriers: It first circulates as large, negatively charged nanoparticles to prolong plasma half-life; then converts to small, positively charged particles via tumor-overexpressed heparanase for deep tumor penetration; and finally assembles into long nanofibers triggered by lysosomal cathepsin B and acidity to extend tumor retention. Importantly, the nanofibers mechanically disrupt lysosomes, increasing lysosomal membrane permeability, inhibiting AKT activation, reducing autophagy, and impairing cytoskeletal integrity─synergistically sensitizing tumors to radiation. This strategy combined with 6 Gy radiation achieved 82.5% tumor suppression in conventional U251 models and 60.4% in radioresistant U87 models, significantly outperforming the clinical radiosensitizer sodium glycididazole. This strategy provides a paradigm for overcoming GBM radioresistance by leveraging bioresponsive nanoscale transformations and lysosomal targeting.

Tanshinone IIA sodium sulfonate (TSS) is a water-soluble derivative of tanshinone IIA, which is the main pharmacologically active component of Salvia miltiorrhiza. This study aimed to verify the preventive and therapeutic effects of TSS and its combined therapeutic effects with magnesium isoglycyrrhizinate (MI) in D-galactosamine- (D-Gal-) induced acute liver injury (ALI) in mice. The potential regulatory mechanisms of TSS on ALI were also examined. Our results may provide a basis for the development of novel therapeutics for ALI.

Stimulation of Hedgehog (Hh) signaling induces carcinogenesis or promotes cell survival in cancers of multiple organs. In epithelial cancer with aberrant Hedgehog activation, abrogation of Hedgehog signaling by cyclopamine, a naturally occurring Hedgehog-specific small-molecule inhibitor, causes profound inhibition of tumor growth. In the present study, cyclopamine displayed a significant potency in suppressing the proliferation of both estrogen-responsive (MCF-7) and estrogen-independent (MDA-MB-231) human breast cancer cells. Cyclopamine induced a robust G1 cell cycle arrest and elicited notable effects on the expression of cyclin D1 through modulation of the MAPK/ERK signaling pathway. Cyclopamine also inhibited the invasive ability of both breast cancer cell lines by suppressing the expression levels of NF-κB, MMP2 and MMP9 protein. Furthermore, in estrogen-responsive MCF-7 cells, cyclopamine significantly downregulated the production of estrogen receptor-α protein. Our results implicate cyclopamine as a novel, potent inhibitor of human breast cancer proliferation and estrogen responsiveness that could potentially be developed into a promising therapeutic agent for the treatment of breast cancer.

The aim of this study was to investigate the effects and mechanisms of antiproliferative transducer of erbB2, 1 (TOB1) on the radiosensitivity of the normal human bronchial epithelial cell line HBE. After exposure to different doses of irradiation or a certain dose for different time intervals, the expression of TOB1 mRNA and protein in HBE cells was determined by semi-quantitative RT-PCR and western blot analysis. Liposome-induced recombinant plasmid transfection and G418 selection were performed to establish a stably transfected TOB1-overexpressing HBE cell line. A clonogenic assay was used to determine the radiosensitivity of the HBE cells with different TOB1 expression statuses. The cell cycle distribution was detected by flow cytometry. The ionizing radiation (IR)-induced γ-H2AX foci formation was detected by immunofluorescence assay. The related mechanism was explored by western blot analysis. TOB1 expression in the HBE cells was not induced by IR, neither dose-dependently nor time-dependently. Compared to the parental or 'mock' transfected HBE cells, the radiosensitivity of HBE cells overexpressing TOB1 was significantly decreased (P<0.05). Exogenous TOB1 prevented HBE cells from apoptosis after IR, in contrast to the control cells (P<0.05), and significantly decreased the IR-induced γ-H2AX foci formation. After IR, the expression of DNA damage repair proteins such as XRCC1, MRE11, FEN1 and ATM was increased in the TOB1‑overexpressing HBE cells when compared with the expression levels in the control cells. HBE/TOB1 cells presented a much higher phosphorylated ERK1/2 and phosphorylated p53 when compared with the levels in the control cell lines when receiving 6 Gy of X-rays. Notably, the increased expression of phosphorylated p53 in HBE/TOB1 cells after IR was sufficiently blocked by U0126, a specific inhibitor of MEK1/2. Different from its functions in several lung cancer cell lines, TOB1 demonstrated a radioprotective function in the immortalized normal human bronchial epithelial cell line HBE via the MAPK/ERK signaling pathway.

Pancreatic cancer is an aggressive malignancy with a characteristic metastatic course of disease and resistance to conventional radiotherapy. As a result, the continual development of novel therapeutic agents is required to improve the current situation. In the present study, the effect of the hedgehog pathway inhibitor, cyclopamine, on cellular radiosensitivity was determined in K‑RASwt Colo‑357 and K‑RASmt SW‑1990 human pancreatic cancer cell lines using the clonogenic survival assay. Apoptosis and cell cycle distribution were detected using flow cytometry assay. Following irradiation (30 mins), residual double‑strand breaks were quantified by identification of γ‑H2AX foci of micronuclei and radiation‑induced γ‑H2AX, p‑ATM, DNA‑PKcs and Ku70 expression was analyzed using western blot analysis. The epidermal growth factor (EGF) and EGF receptor (EGFR) inhibitor, gefitinib, were utilized to determine the related mechanisms. The results revealed that cyclopamine treatment significantly reduced cell clonogenic survival but failed to induce apoptosis and radiation‑induced G2 arrest. Flow cytometry revealed that cyclopamine treatment enhanced γ‑H2AX foci in Colo‑357 and SW‑1990 cells exposed to irradiation. In addition, radiation‑induced p‑ATM, DNA‑PKcs and Ku70 were all inhibited. EGF also rescued pancreatic cancer cells from cyclopamine‑induced H2AX phosphorylation following irradiation. Thus, cyclopamine enhanced the radiosensitivity of human pancreatic cancer cells, in part, through an EGFR‑dependent pathway, indicating a rational approach in combination with radiotherapy.

Objective To study the impacts of berberine on the growth, migration and radiosensitivity in human breast cancer cells.Methods MTT assay was used to evaluate cell growth.In vitro scratch migration assay was used to determine cell migration.Annexin V assay was used to detect cell apoptosis.The distribution of cell cycle was evaluated by flow cytometry assay.Colony formation assay was used to detect the influence of berberine on cell radiosensitivity. Western blot assay was employed to measure protein expression.Results Berberine inhibited cell growth and migration in two human breast cancer cell lines, MCF-7 and MDA-MB-231, in a dose-and time-dependent manner. Furthermore,berberine resulted in a cell cycle G0/G1 arrest.Compared with control,the early apoptosis in MDA-MB-231 and MCF-7 cells treated with 40 pμmol/L of berberine was as high as 86.6％ and 66.6％ (t =8.79,10.32,P ＜ 0.01 ),respectively. Berberine caused a dose-dependent increase in Bax and Caspase-3 protein expressions,but did not change Cyclin D1 protein expression,while suppressed the expressions of Cyclin B1 and Bcl-2 protein. As analyzed with multi-target click model fitting curves,the SERD0 of berberine-treated cells were 1.12 and 1.22 for MDA-MB-231 and MCF-7cells respectively at the dose D0 of X-rays.Conclusions The berberine inhibited the growth and migration of breast cancer cells via apoptosis induction and cell cycle arrest.Moreover,berberine increases cell sensitivity to X-ray irradiation.

Objective To study the impact of cyclopamine on radiosensitivity in lung cancer cell lines and normal lung epithelial cells.Methods MTT assay was used to determine cell growth and survival;clonogenic assay was employed to detect the impact of cyclopamine on the radiosensitivity of lung cancer cells;Western blot assay was employed to measure protein expression;Shh-siRNA was transfected into NIH-H226 cells to observe the influence of the Shh expression decrease on the radiosensitivity.Results 2.5 μmol/L of cyclopamine showed no significant influence on the growth and proliferation of both lung cancer cells and normal lung epithelial cells.However,the dose of 2.5 μmol/L significantly increased radiosensitivity of five human lung cancer cell lines,including NIH-H520,NIH-H1437,NIH-H1975,A549 and NIH-H226,but failed to affect radiosenstivity of three normal lung epithelial cell lines,including IMR90,CCD-16Lu and BEAS-2B.After the transfection of Shh-siRNA,the radiosensitivity of NIH-H226 cells was enhanced obviously.Conclusion Cyclopamine significantly reduced Shh protein expression in five lung cancer cell lines.A significant reducation of Shh protein by Shh-siRNA increased susceptibility of lung cancer cells to radiation and disrupted cyclopamine-mediated radiosenstivity.The present findings demonstrated for the first time that cyclopamine selectively increases radiosenstivity of lung cancer cells,the alteration of Shh expression could be one of the mechanisms involved in cyclopamine-mediated radiosenstivity.

Objective To study and compare the local control rate of the primary location and lymph node metastasis for nasopharyngeal carcinoma between intensitymodulated radiation therapy (IMRT) and conventional radiation therapy (CRT).Methods Thirty nasopharyngeal carcinoma patients in stage T3N1M0 planned with CMS system in IMRT and CRT,calculated the dose to tumor and normal tissues respectively.The 2-year and 3-year control rate was summarized.Results There was no significant difference about the dose to tumor between IMRT and CRT,but the dose to the normal tissuew was lower in IMRT than in CRT.The 2-year and 3-year control rate were 95%,93% with IMRT and 83%,75% with CRT.Conclusion The dose distribution to tumor target area is more creditable with IMRT than with CRT.The dose to the normal tissues is controlles effectively with IMRT,IMRT is more advantageous than CRT.

Objective: To investigate the effect of cholesteryl ester transfer protein (CETP) on the radiosensitivity of human cervical cancer HeLa cells. Methods: HeLa-CETP cells with overexpression of CETP were established by stable transfection with pcDNA3 expression vector encoding a full-length of CETP gene. The HeLa cells which transfected with pcDNA3 empty vector were used as the control. The survival rate of HeLa-CETP cells was examined by clone formation assay. The apoptosis rate was detected by FCM. The expressions of apoptosis-related proteins Bax, Bcl-2 and nuclear factor-kappa B (NF-κB) were detected by Western blotting. Results: After X-ray radiation exposure, the sensitizing enhancement ratio (SER)D0 and SERDq of HeLa-CETP cells with overexpression of CETP were 1.42 and 2.09, respectively. There was no significant difference in the survival rate between the untransfected HeLa cells and the HeLa cells transfected with empty vector. FCM result showed that X-ray-induced apoptosis of HeLa-CETP cells was significantly higher than those of the untransfected HeLa cells and the HeLa cells transfected with empty vector (P＜0.05). Moreover, the enhanced expression level of CETP could increase the expression of pro-apoptosis-related Bax protein and decrease the expressions of anti-apoptosis-related Bcl-2 and NF-кB proteins. Conclusion: Elevated expression of CETP by transfection with exogenous CETP gene can increase the radiation sensitivity of HeLa cells. This mechanism may be related to the increased radiation-induced apoptosis and the changes in the expression of apoptosis-related proteins.. DOI:10.3781/j.issn.1000-7431.2011.07.007

In order to investigate the effects of human anti-proliferative protein Tob1 (transducer of ErbB2,1) on the radio-sensitivity of human cervix cancer cells HeLa,the full length Tob1 recombinant plasmid pcDNA3.0/Tob1 (pc3/Tob1) was constructed and transfected into HeLa cells by lipofectamine. And G418 selection was used to get HeLa cell line stably expressed exogenous Tob1. The clonogenic assay was applied to study the effects of Tob1 on HeLa cell radio-sensitivity,while flow cytometry assay and Western Blot analysis were performed to determine the related mechanisms. It was shown that,compared with parental HeLa cells and mock plasmid pcDNA3.0 transfected HeLa/pc3,the radio-sensitivity of HeLa cells transfected with Tob1 was increased obviously. It was also found that increased Tob1 expression could enhance cell apoptosis of HeLa induced by irradiation,while up-regulated protein expression level of Bax,but down-regulated Bcl2 expression occur at the same time. These results suggested that Tob1 might play a role as radio-sensitizer of cervix cancer cell line HeLa via regulating the expression of apoptosis related genes.

Objective To study the anti-cancer effects of sanguinarine using two human cervical cancer HeLa and Siha cell lines.Methods MTr and colony formation were used to observe cell growth.Cell scratch test was performed to evaluate cell migration and invasion.Results After treatment with 1.0μmol/L and 5.0μmol/L SAN,the viability of Siha cells was 72%and 10%(P＜0.01),respectively.HeLa cells were more sensitive than Siha cells.After treatment with 1.0 μmol/L SAN,colony formation in HeLa and Siha cells decreased from 33 to 14 and 16 (P＜0.05).Cell scratch test showed cut width was(0.5 1±0.04)mm and(0.64±0.02)mm in the control and(1.22±0.02)mm and (1.63±0.01)mm in the HeLa and Siha cells treated with 0.5 μmol/L SAN(P＜0.01 and P＜0.05).MTT,colony formation and cell scratch test showed time-or dose-dependent manner.Conclusion SAN could inhibit the proliferation,migration and invasion of HeLa and Siha cells,which provide a laboratory evidence for potential application of SAN in clinical therapy of cervical cancer.