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      • IGF-1 from Adipose-Derived Mesenchymal Stem Cells Promotes Radioresistance of Breast Cancer Cells

        Yang, Hui-Ying,Qu, Rong-Mei,Lin, Xiao-Shan,Liu, Tong-Xin,Sun, Quan-Quan,Yang, Chun,Li, Xiao-Hong,Lu, Wei,Hu, Xiao-Fang,Dai, Jing-Xing,Yuan, Lin Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.23

        Purpose: The aim of this study was to investigate effects of adipose-derived mesenchymal stem cells (AMSCs) on radioresistance of breast cancer cells. Materials and Methods: MTT assays were used to detect any influence of AMSC supernatants on proliferation of breast cancer cells; cell migration assays were used to determine the effect of breast cancer cells on the recruitment of AMSCs; the cell survival fraction post-irradiation was assessed by clonogenic survival assay; ${\gamma}$-H2AX foci number post-irradiation was determined via fluorescence microscopy; and expression of IGF-1R was detected by Western blotting. Results: AMSC supernatants promoted proliferation and radioresistance of breast cancer cells. Breast cancer cells could recruit AMSCs, especially after irradiation. IGF-1 derived from AMSCs might be responsible for the radioresistance of breast cancer cells. Conclusions: Our results suggest that AMSCs in the tumor microenvironment may affect the outcome of radiotherapy for breast cancer in vitro.

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        Generation of Buffalo Mammary Epithelial Cells with Targeted Knockout of 18s rDNA by a CRISPR/Cas9 Adenovirus System

        Peng Zhu,An-Qin Duan,Ting-Xian Deng,Xing-Rong Lu,Xiao-Ya Ma,Sha-Sha Liang,Chun-Ying Pang,Xian-Wei Liang 한국생물공학회 2017 Biotechnology and Bioprocess Engineering Vol.22 No.5

        In recent years, CRISPR/Cas9 has rapidly become one of the most promising genome editing tools because it is simple and easy to use and cost effective. However, the large size of Cas9 sequences limits its application in clinically promising vectors and it also impacts non-viral transfection. In this study, CRISPR/Cas9 adenovirus vectors that target the buffalo 18s rDNA gene were constructed, transfected into 293 cells for adenovirus packaging, and the adenovirus was used to knockout the 18s rDNA gene in buffalo mammary epithelial cells. The results demonstrated that the CRISPR/Cas9 adenovirus vectors for the buffalo 18s rDNA gene could efficiently target the sites as revealed by the fluorescence reporter system. After amplification, the adenovirus titer of Sn458- 18s1 and Sn458-18s2 reached 1.03 × 109PFU/mL and 1.05 × 109 PFU/mL, respectively. For buffalo mammary epithelial cell infection, the efficiency was 100% when the multiplicity of infection (MOI) 100 PFU/mL. There were 9 mutational clones found in the 20 clones, and the gene mutagenesis rate reached 45%. Of these, 2 clones were 35-bp deleted and 7 clones were 12-bp deleted. These results suggested that the adenovirus system overcame the low transfection efficiency of the buffalo mammary epithelial cells associated with using lipid-based methods or electroporation. Moreover, we preliminary developed an efficient technique for multiple-locus gene targeting at repeated sequences of the buffalo genome.

      • Analysis of Prethrombotic States in Patients with Malignant Tumors

        Cui, Lin,Sun, You-Hong,Chen, Jue,Wang, Lu,Liu, Jian-Jun,Zhou, Xiang-Rong,Ding, Jie,Liu, Xing-Xiang,Huang, Xin-En Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.13

        Background: This study aimed to investigate the incidence and risk factors for a prethrombotic state in patients with malignant tumors. Materials and Methods: Plasma d-dimer (D-D) in patients with malignant tumors was measured. Abnormal rates of D-D and possible risk factors like gender, age, type of tumor, and staging of tumor were analyzed. Results: Of 1,453 patients, 629 demonstrated plasma D-D abnormality (43.3%). The D-D abnormal rate of male patients (n=851, 43.5%) was not statistically significantly different from that for female patients (n=602, 43.0%) (p>0.05). D-D abnormal rate increased with age and was statistically significant among different age groups (p<0.05). Regarding staging of tumor, D-D abnormal rate in patients with phase I was 2.0%, 6.2% in phase II, 47.6% in phase III and 83.1% in phase IV, with statistically significant differences between phase III and II, as well as phase III and IV (p<0.01). Conclusions: A prethrombotic state was closely related to malignancy of tumors. The risk factors for a prethrombotic state include age and tumor stage.

      • Establishment of Paclitaxel-resistant Breast Cancer Cell Line and Nude Mice Models, and Underlying Multidrug Resistance Mechanisms in Vitro and in Vivo

        Chen, Si-Ying,Hu, Sa-Sa,Dong, Qian,Cai, Jiang-Xia,Zhang, Wei-Peng,Sun, Jin-Yao,Wang, Tao-Tao,Xie, Jiao,He, Hai-Rong,Xing, Jian-Feng,Lu, Jun,Dong, Ya-Lin Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.10

        Background: Breast cancer is a common malignant tumor which affects health of women and multidrug resistance (MDR) is one of the main factors leading to failure of chemotherapy. This study was conducted to establish paclitaxel-resistant breast cancer cell line and nude mice models to explore underlying mechanisms of MDR. Methods: The breast cancer drug-sensitive cell line MCF-7 (MCF-7/S) was exposed in stepwise escalating paclitaxel (TAX) to induce a resistant cell line MCF-7/TAX. Cell sensitivity to drugs and growth curves were measured by MTT assay. Changes of cell morphology and ultrastructure were examined by optical and electron microscopy. The cell cycle distribution was determined by flow cytometry. Furthermore, expression of proteins related to breast cancer occurrence and MDR was tested by immunocytochemistry. In Vivo, nude mice were injected with MCF-7/S and MCF-7/TAX cells and weights and tumor sizes were observed after paclitaxel treatment. In addition, proteins involved breast cancer and MDR were detected by immunohistochemistry. Results: Compared to MCF-7/S, MCF-7/TAX cells had a higher resistance to paclitaxel, cross-resistance and prolonged doubling time. Moreover, MCF-7/TAX showed obvious alterations of ultrastructure. Estrogen receptor (ER) expression was low in drug resistant cells and tumors while expression of human epidermal growth factor receptor 2 (HER2) and Ki-67 was up-regulated. P-glycoprotein (P-gp), lung resistance-related protein (LRP) and glutathione-S-transferase-${\pi}$ (GST-${\pi}$) involved in the MDR phenotype of resistant cells and tumors were all overexpressed. Conclusion: The underlying MDR mechanism of breast cancer may involve increased expression of P-gp, LRP and GST-${\pi}$.

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