泛素E3连接酶MARCH5对子宫颈癌细胞增殖、侵袭和迁移的影响

Role of E3 ubiquitin ligase MARCH5 in proliferation,invasion and migration of cervical cancer cells

目的 探讨泛素E3连接酶膜相关锌指蛋白5 （membrane-associated RING-CH 5,MARCH5）在子宫颈癌组织中的表达、功能以及潜在机制。方法 用免疫组织化学检测泛素E3连接酶MARCH5在宫颈癌组织中的表达,EdU、Transwell实验和划痕实验检测沉默泛素E3连接酶MARCH5后,宫颈癌细胞增殖、侵袭和迁移能力变化情况,Western blot检测宫颈癌HeLa细胞株沉默MARCH5基因后Drp1、Bax、MARCH5、PGAM的表达情况。使用Mito Tracker Green标记线粒体,然后使用免疫荧光检测MARCH5与线粒体的共定位。结果 在宫颈癌组织中MARCH5的表达明显较正常宫颈组织高,且MARCH5的表达与宫颈癌的分级有关（P 〈0. 01）,与年龄和分期无关（P〉 0. 05）。沉默MARCH5后,HeLa细胞的增殖、侵袭和迁移能力降低（P 〈0. 05）。沉默MARCH5后,Drp1表达下调,PGAM5和Bax表达上调（P 〈0. 05）。结论 MARCH5在宫颈癌组织中表达上调,是宫颈癌候选癌基因,也是宫颈癌治疗的一个潜在靶点。

Objective To study the expression of membrane-associated RING-CH 5 （MARCH5）, an E3 ubiquitin ligase, in cervical cancer and investigate its role in the proliferation, invasion and migration of cervical cancer cells in vitro. Methods Immunohistochemistry was used to detect the expression of MARCH5 in clinical specimens of cervical cancer tissues. The cervical cancer cell line HeLa with MARCH5 silencing was examined for changes in proliferation, invasion, and migration abilities using EdU assay, Transwell assay, and scratch wound healing assay, and Western blotting was used to detect the expression of Drpl, Bax, MARCH5, and PGAM in the ceils. Immunofluorescence assay with MitoTracker labeling was employed to characterize the colocalization of MARCH5 with the mitochondria. Results Cervical cancer tissues expressed high levels of MARCH5 compared with the normal cervical tissues, and the expression level was correlated with the tumor grade （P 〈 0.01 ） but not with the patients＇ age or tumor stage （P 〉 0.05）. Silencing MARCH5 in HeLa cells resulted in significant suppression of the cell proliferation, invasion, and migration （P 〈 0.05） , and significantly down-regulated Drpl expression and up-regulated PGAM5 and Bax expression （P 〈 0.05 ）. Conclusion oncogene for cervical cancer MARCH5, which is up-regulated in cervical cancer, can serve as a candidate and also a potential target for cervical cancer treatment.