Objective To observe the influence of trastuzumab on DNA break repair and Her-2 nuclear import after radiation in breast cancer cell line SKBR3,and discuss the radiosensitivity mechanism of trastuzumab.Methods Clone formation assay was used to analyze the difference of survival fractions between radiation group and radiation plus trastuzumab group.Confocal microscopy was applied to observe the influence of trastuzumab in the nuclear import process of Her-2 and the expression of γH2AX after radiation,which is considered as the marker of DNA double strand break.Western blotting was used to detect the expression of Her-2 and DNA-PKcs in nuclei after radiation.Results The result of clone formation assayshowed that the SF2 in radiation group was 0.547±0.046 and 0.321±0.022 in the radiation plus trastuzumab group were significantly decreased,the results of confocal microscopy showed that trastuzumab postponed the nuclear import process of Her-2 (52.80±19.74 in radiation group,21.41±10.55 in the radiation group),and increased expression of γH2AX after radiation (85.40±25.63 in radiation group,18.53±44.32 in the radiation group),and western blotting revealed trastuzumab reduced the expression of Her-2,DNA-PKcs in nuclei.Conclusion Trastuzumab can inhibit the radiation induced nuclear import of Her-2,and decrease Her-2,DNA-PKcs in nuclei to increase the DSB on early stage after radiation.

This study examined the radiation-induced ERBB2 nuclear transport in the BT474 breast cancer cell line and the relationship between caveolin-1 and radiation-induced ERBB2 nuclear transport. The BT474 cells were treated with herceptin (200 nmol/L), PP2 (a caveolin-1 inhibitor, 100 nmol/L) and irradiation combined or alone. Confocal microscopy was used to observe the nuclear import of ERBB2 and caveolin-1 after irradiation. Western blotting was employed to detect the expression of ERBB2, caveolin-1 and DNA-PKcs after irradiation, and immunoprecipitation to identify the ERBB2 and caveolin-1 complex before perinuclear ERBB2 localization. Confocal microscopy showed the transport of ERBB2 and caveolin-1 from the cell membrane to the nucleus 15 min after irradiation and the proteins accumulated at the perinuclear region within 45 min. Western blotting revealed that the expression levels of ERBB2, caveolin-1 and DNA-PKcs were increased after irradiation and reached a peak 45 min later. Both herceptin and PP2 treatments were found to decrease ERBB2 expression. An immune complex composed of ERBB2 and caveolin-1 was found in the herceptin group after irradiation. It was concluded that after irradiation, ERBB2 may be transported from the cell membrane to the nucleus and activate DNA-PKcs to trigger DNA double-strand break (DSB) repair; caveolin-1 may participate in this process. Treatments involving the downregulation of caveolin-1 may increase the radiosensitization of breast cancer cells.

This study examined the radiation-induced ERBB2 nuclear transport in the BT474 breast cancer cell line and the relationship between caveolin-1 and radiation-induced ERBB2 nuclear transport. The BT474 cells were treated with herceptin (200 nmol/L), PP2 (a caveolin-1 inhibitor, 100 nmol/L) and irradiation combined or alone. Confocal microscopy was used to observe the nuclear import of ERBB2 and caveolin-1 after irradiation. Western blotting was employed to detect the expression of ERBB2, caveolin-1 and DNA-PKcs after irradiation, and immunoprecipitation to identify the ERBB2 and caveolin-1 complex before perinuclear ERBB2 localization. Confocal microscopy showed the transport of ERBB2 and caveolin-1 from the cell membrane to the nucleus 15 min after irradiation and the proteins accumulated at the perinuclear region within 45 min. Western blotting revealed that the expression levels of ERBB2, caveolin-1 and DNA-PKcs were increased after irradiation and reached a peak 45 min later. Both herceptin and PP2 treatments were found to decrease ERBB2 expression. An immune complex composed of ERBB2 and caveolin-1 was found in the herceptin group after irradiation. It was concluded that after irradiation, ERBB2 may be transported from the cell membrane to the nucleus and activate DNA-PKcs to trigger DNA double-strand break (DSB) repair; caveolin-1 may participate in this process. Treatments involving the downregulation of caveolin-1 may increase the radiosensitization of breast cancer cells.
Active Transport, Cell Nucleus ; physiology ; Breast Neoplasms ; metabolism ; physiopathology ; Caveolin 1 ; metabolism ; Cell Line, Tumor ; Female ; Humans ; Protein Transport ; physiology ; Radiation ; Receptor, ErbB-2 ; metabolism

Objective To analyze the setup errors by cone-beam computed tomography (CBCT) for breast cancer patients who were immobilized with neck and breast thermoplastic mask and received intensity modulated radiation therapy (IMRT), and to calculate the external margins from the clinical target volume (CTV) to the planning target volume (PTV) (MPTV) of tumors. Methods Twenty-five breast cancer patients who were immobilized with neck and breast thermoplastic mask and received IMRT in the Oncology Department of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from November 2016 to June 2017 were enrolled. The position of the patients were verified by CBCT before treatment . The linear and rotation errors of the X, Y and Z axes were analyzed by online bone registration. The systematic errors (Σ) and random errors (σ) of the patients were also calculated, and then the margins from CTV to PTV margins were calculated based on MPTV=2.5Σ+0.7σ. 25 patients'height, weight, body mass index (BMI) and the maximum diameters of CTV in the lateral, longitudinal and vertical directions were recorded, and the relation between the setup errors and the above mentioned was analyzed by using Spearman method. Results A total of 174 CBCT scans for 25 breast cancer patients were completed. The group Σ were 1.40 mm, 1.50 mm and 1.20 mm, and rotation errors were 0.9°, 0.7° and 0.8° at the X, Y and Z axes, respectively. The group σ were 2.20 mm, 3.00 mm and 1.40 mm, and rotation errors were 0.7°, 0.6° and 0.7° at the X, Y and Z axes, respectively. MPTVwas recommended as 4.90 mm, 6.00 mm and 3.90 mm at the X, Y and Z axes, respectively. There was no correlation between the height, weight, BMI of the patients and the setup errors (all P ＞ 0.05). However, there was a significant correlation between the maximum lateral, longitudinal diameters of the CTV and the setup errors (rs= 0.406, P= 0.044; rs= 0.512, P= 0.009). Conclusions The neck and breast thermoplastic mask can improve the diagnostic accuracy of radiotherapy in breast cancer patients. The data of setup errors verified by CBCT can provide meaningful references for the setting of MPTV.

Objective To investigate the accuracy and application value of optical surface monitoring system in intensity modulated radiotherapy for thoracic tumors patients.Methods Twenty-eight patients with thoracic tumors were included.During each treatment fraction,the patients were immobilized with body surface markers and laser lamps.The surface images obtained by the optical surface monitoring system were registered with the reference images and recorded during the CBCT scan.The translation and rotation errors of x (left-right),y (craniocaudal) and z (anterior-posterior) axes were recorded.After scanning,the CBCT images were registered with the planned CT images and the translation and rotation errors of x,y and z axes were recorded.The setup errors of these two image systems were analyzed and corrected before each treatment.The correlation between the two sets of setup errors were analyzed with Pearson test,and systematic error (∑) and random error (σy) were also calculated.The consistency of the two image systems was evaluated with the Bland-Altman method and the 95％ limits of agreement were calculated.Results There was a good correlation between these two groups,and the correlation coefficients were 0.79,0.62,and 0.53 in x,y and z axes,respectively.The ∑/σr of the optical surface monitoring system were 0.7 mm/1.5 mm,0.9 mm/1.8 mm and 0.9 mm/1.5 mm in x,y and z axes,respectively.The ∑/σ of CBCT were 0.8 mm/1.6 mm,1.3 mm/1.9 mm and 0.7 mm/1.5 mm in x,y and z axes,respectively.The 95％ limits of agreement of translations direction were (-2.0-2.3),(-3.4-3.6) and (-3.3-2.4) mm,and the 95％ limits of agreement of rotation direction were (-2.0 to 1.6)°,(-2.0 to 1.4)° and (-1.6 to 1.6)° inx,y and z axes,respectively.Conclusions The optical surface monitoring system is an effective image guide tool,which can quickly and accurately verify the patient's position and improve the position accuracy.It can be applied for positioning in the intensity modulated radiation treatments for the thoracic tumor patients.

Background::Although some well-established oncogenes are involved in cancer initiation and progression such as prostate cancer (PCa), the long tail of cancer genes remains to be defined. Goosecoid ( GSC) has been implicated in cancer development. However, the comprehensive biological role of GSC in pan-cancer, specifically in PCa, remains unexplored. The aim of this study was to investigate the role of GSC in PCa development. Methods::We performed a systematic bioinformatics exploration of GSC using datasets from The Cancer Genome Atlas, Genotype-Tissue Expression, Gene Expression Omnibus, German Cancer Research Center, and our in-house cohorts. First, we evaluated the expression of GSC and its association with patient prognosis, and identified GSC-relevant genetic alterations in cancers. Further, we focused on the clinical characterization and prognostic analysis of GSC in PCa. To understand the transcriptional regulation of GSC by E2F transcription factor 1 ( E2F1), we performed chromatin immunoprecipitation quantitative polymerase chain reaction (qPCR). Functional experiments were conducted to validate the effect of GSC on the tumor cellular phenotype and sensitivity to trametinib. Results::GSC expression was elevated in various tumors and significantly correlated with patient prognosis. The alterations of GSC contribute to the progression of various tumors especially in PCa. Patients with PCa and high GSC expression exhibited worse progression-free survival and biochemical recurrence outcomes. Further, GSC upregulation in patients with PCa was mostly accompanied with higher Gleason score, advanced tumor stage, lymph node metastasis, and elevated prostate-specific antigen (PSA) levels. Mechanistically, the transcription factor, E2F1, stimulates GSC by binding to its promoter region. Detailed experiments further demonstrated that GSC acted as an oncogene and influenced the response of PCa cells to trametinib treatment. Conclusions::GSC was highly overexpressed and strongly correlated with patient prognosis in PCa. We found that GSC, regulated by E2F1, acted as an oncogene and impeded the therapeutic efficacy of trametinib in PCa.

In fixed prosthodontics, clear exposure of the preparation margin is the prerequisite for obtaining accurate digital impressions and improving the marginal fit of restorations. To resolve the issues associated with the cord retraction technique, such as pain, acute injury, and prolonged procedural time, this study proposes a new technology for intraoral digital impression taking with pneumatic gingival retraction. The new scanning head blows a high-speed airflow that instantaneously separates the free gingiva, locally exposing the subgingival preparation margin. Combined with the farthest point preservation stitching algorithm based on the distance from the normal vector and high-speed laser scanning photography, it achieves global preparation edge data and gingival reconstruction, realizing painless, non-invasive, and efficient precise acquisition of the preparation margin. Using this new technique, a patient with a full porcelain crown restoration on a posterior tooth was treated. The digital impression revealed a clear margin of the preparation, and the crown made from this data has a good marginal fit.