Ovarian cancer(OC)is the sixth most common cancer and the seventh cause of death from cancer in women.The etiology and the ovarian carcinogenesis still need clarification although ovulation may be determinant due to i...Ovarian cancer(OC)is the sixth most common cancer and the seventh cause of death from cancer in women.The etiology and the ovarian carcinogenesis still need clarification although ovulation may be determinant due to its carcinogenic role in ovarian surface epithelium.The link between ovarian carcinogenesis and DNA repair is well established and it became clear that alterations in DNA damage response may affect the risk to develop OC.Polymorphisms are variations in the DNA sequence that exist in normal individuals of a population and are capable to change,among other mechanisms,the balance between DNA damage and cellular response.Consequently,genetic variability of the host has a great role in the development,progression and consequent prognosis of the oncologic patient as well as in treatment response.Standard treatment for OC patients is based on cytoreductive surgery,followed by chemotherapy with a platinum agent and a taxane.Although 80%of the patients respond to the first-line therapy,the development of resistance is common although the mechanisms underlying therapy failure remain mostly unknown.Because of their role in oncology,enzymes involved in the DNA repair pathways,like DNA Ligase IV(LIG4),became attractive study targets.It has been reported that variations in LIG4 activity can lead to a hyper-sensitivity to DNA damage,deregulation of repair and apoptosis mechanisms,affecting the susceptibility to cancer development and therapy response.To overcome resistance mechanisms,several investigations have been made and the strategy to target crucial molecular pathways,such as DNA repair,became one of the important areas in clinical oncology.This review aims to elucidate the link between DNA repair and OC,namely which concerns the role of LIG4 enzyme,and how genetic polymorphisms in LIG4 gene can modulate the activity of the enzyme and affect the ovarian carcinogenesis and treatment response.Moreover,we try to understand how LIG4 inhibition can be a potential contributor for the development of new cancer treatm展开更多
为了筛选靶向DNA连接酶Ⅳ的抑制剂以实现更有效的基因定点插入,采用分子对接技术筛选出与前期研究中靶向DNA连接酶Ⅳ的抑制剂SCR7作用类似的小分子化合物。筛选到与化合物库一致的nocodazole、Fisetin和Methylene blue 3个小分子化合物...为了筛选靶向DNA连接酶Ⅳ的抑制剂以实现更有效的基因定点插入,采用分子对接技术筛选出与前期研究中靶向DNA连接酶Ⅳ的抑制剂SCR7作用类似的小分子化合物。筛选到与化合物库一致的nocodazole、Fisetin和Methylene blue 3个小分子化合物,通过用MSTN基因T11位点序列截断的Firefly Luciferase荧光素酶报告载体结合CRISPR/Cas9-gRNA-T11载体共转细胞,并结合不同的小分子化合物处理。结果表明,没有用小分子化合物处理时,萤火虫荧光素酶被截断的位置发生NHEJ修复,不能得到大量有活性的萤火虫荧光素酶;经过小分子化合物处理,抑制了细胞内的NHEJ,提高了HDR效率,能得到大量有活性的萤火虫荧光素酶。使用nocodazole、Methylene blue和Fisetin处理后的萤火虫荧光素酶检测结果65256.3、53713和77058.3分别是SCR7处理后的萤火虫荧光素酶检测结果41905.3的1.6、1.3和1.8倍,是没有SCR7处理后的萤火虫荧光素酶检测结果10120的6.4、5.3和7.6倍。证明所用的筛选策略正确,所筛选出的小分子化合物是具有DNA ligaseⅣ抑制活性的抑制剂,为后续研究奠定了基础。展开更多
基金Supported by Research Department of Portuguese League against Cancer(NRNorte)and Minister of Health of Portugal(CFICS-45/2007)
文摘Ovarian cancer(OC)is the sixth most common cancer and the seventh cause of death from cancer in women.The etiology and the ovarian carcinogenesis still need clarification although ovulation may be determinant due to its carcinogenic role in ovarian surface epithelium.The link between ovarian carcinogenesis and DNA repair is well established and it became clear that alterations in DNA damage response may affect the risk to develop OC.Polymorphisms are variations in the DNA sequence that exist in normal individuals of a population and are capable to change,among other mechanisms,the balance between DNA damage and cellular response.Consequently,genetic variability of the host has a great role in the development,progression and consequent prognosis of the oncologic patient as well as in treatment response.Standard treatment for OC patients is based on cytoreductive surgery,followed by chemotherapy with a platinum agent and a taxane.Although 80%of the patients respond to the first-line therapy,the development of resistance is common although the mechanisms underlying therapy failure remain mostly unknown.Because of their role in oncology,enzymes involved in the DNA repair pathways,like DNA Ligase IV(LIG4),became attractive study targets.It has been reported that variations in LIG4 activity can lead to a hyper-sensitivity to DNA damage,deregulation of repair and apoptosis mechanisms,affecting the susceptibility to cancer development and therapy response.To overcome resistance mechanisms,several investigations have been made and the strategy to target crucial molecular pathways,such as DNA repair,became one of the important areas in clinical oncology.This review aims to elucidate the link between DNA repair and OC,namely which concerns the role of LIG4 enzyme,and how genetic polymorphisms in LIG4 gene can modulate the activity of the enzyme and affect the ovarian carcinogenesis and treatment response.Moreover,we try to understand how LIG4 inhibition can be a potential contributor for the development of new cancer treatm
