卵巢癌组织中PLOD2的表达及其在缺氧条件下通过PI3K/Akt通路对卵巢癌细胞作用的研究

目的探究前胶原-赖氨酸,2-氧戊二酸5-双加氧酶2(PLOD2)对卵巢癌细胞的作用及其可能的作用机制。方法收集2017年8月至2019年1月在海南医学院附属海南医院行手术治疗的33例卵巢癌患者的癌组织标本作为卵巢癌组,另收集同期的行良性卵巢囊肿切除33例患者的正常卵巢组织作为正常组。采用实时荧光定量聚合酶链反应(RT-PCR)?免疫组化(IHC)和Western Blot检测组织中PLOD2的表达;将卵巢癌细胞SK-OV-3进行缺氧处理,采用RT-PCR检测PLOD2 mRNA的表达;采用Western Blot检测PLOD2?p-PI3 K?PI3 K?p-Akt和Akt的蛋白表达;采用CCK-8实验检测细胞活力;采用5-乙炔基-2′-脱氧尿苷(EdU)实验检测细胞增殖;采用Transwell实验检测细胞迁移和侵袭。结果与正常组比较,卵巢癌组卵巢组织中PLOD2 mRNA和蛋白表达?IHC评分明显升高(P<0.05)。与缺氧0h的细胞比较,缺氧24h和缺氧48h细胞中PLOD2 mRNA表达和蛋白表达明显升高(P<0.05);与缺氧24h的细胞比较,缺氧48h细胞中PLOD2 mRNA表达和蛋白表达明显升高(P<0.05)。在缺氧条件下,与si-NC组相比,si-PLOD2组细胞在24h?48h和72h的细胞活力?EdU+细胞数?迁移细胞数以及侵袭细胞数明显降低(P<0.05),PLOD2?p-PI3 K/PI3 K和p-Akt/Akt表达明显降低(P<0.05)。结论卵巢癌组织中PLOD2高表达,在缺氧条件下,PLOD2可能通过激活PI3 K/Akt通路促进卵巢癌细胞增殖?迁移和侵袭。

The expression of PLOD2 in ovarian cancer tissues and its effect on ovarian cancer cells through PI3K/Akt pathway under hypoxic conditions

Objective To explore the role of procollagen-lysine,2-oxoglutarate 5-dioxygenase 2(PLOD2)in ovarian cancer and its possible mechanism.Methods Cancer tissue samples from 33 patients with ovarian cancer who underwent surgical treatment in Hainan Hospital Affiliated to Hainan Medical College from August 2017 to January 2019 were collected as the ovarian cancer group,and normal ovarian tissue samples from 33 patients who underwent benign ovarian cyst resection during the same period were collected as the normal group.Real time fluorescent polymerase chain reaction(RT-PCR),immunohistochemistry(IHC)and Western Blot were used to detect the expression of PLOD2 in the tissues.Ovarian cancer cells SK-OV-3 were treated with hypoxia,and expression of PLOD2 mRNA was detected by RT-PCR.Western Blot was used to detect expression of PLOD2,p-PI3 K,PI3 K,p-Akt and Akt protein in cells.CCK-8 experiment was used to detect cell viability.EdU experiment was used to detect cell proliferation.Transwell experiment was used to detect cell migration and invasion.Results Compared with the normal group,the expression of PLOD2 mRNA and protein,IHC score in the ovarian cancer group were significantly increased(P<0.05).Compared with the 0h hypoxia group,the PLOD2 mRNA expression and protein expression in the cells of the 24h hypoxia group and the 48h hypoxia group were significantly increased(P<0.05).Compared with the 24h hypoxia group,the expression of PLOD2 mRNA and protein in the cells of the 48h hypoxia group was significantly increased(P<0.05).Under hypoxic conditions,compared with the si-NC group,the cell viability at 24h,48h and 72h,EdU+cell number,migrating cell number and invading cell number of the si-PLOD2 group were significantly reduced(P<0.05),PLOD2,p-PI3 K/PI3 K and p-Akt/Akt expression were significantly reduced(P<0.05).Conclusions PLOD2 is highly expressed in ovarian cancer tissues.Under hypoxia,PLOD2 may promote the proliferation,migration and invasion of ovarian cancer cells by activating the PI3 K/Akt pathway.