C-末端Src蛋白/黏着斑激酶/上皮钙黏素通路在宫颈癌侵袭转移中的作用及机制研究

Study of the role and mechanism of C-terminal Src protein/focal adhesion kinase/E-cadherin pathway in invasion and metastasis of cervical cancer

目的研究C-末端Src蛋白(c-Src)/黏着斑激酶(FAK)/上皮钙黏素(E-cad)通路在宫颈癌侵袭转移中的作用。方法选取来自湖南医药学院第一附属医院2019年1月至2020年12月收集的宫颈鳞癌(CSCC)组织标本30例及正常宫颈(NC)组织标本30例,采用免疫组织化学染色检测CSCC、NC组织中E-cad阳性率,分析E-cad与CSCC患者临床病理特征的关系。采用Western blot检测CSCC、NC组织中c-Src、FAK、E-cad蛋白表达水平。另选取宫颈癌Caski细胞系,将其分为Caski组、Caski+c-Src抑制剂组,Caski组不进行处理,Caski+c-Src抑制剂组加入c-Src抑制剂(2.7 nmol/L),观察两组迁移、侵袭能力的变化及凋亡情况,采用Western blot检测两组c-Src、FAK、E-cad蛋白表达水平。结果CSCC组织标本的E-cad阳性率低于NC组织标本,差异有统计学意义(P<0.05)。E-cad表达与CSCC患者的国际妇产科联盟分期、分化程度、复发情况相关联(P<0.05)。CSCC组织c-Src、FAK蛋白表达水平高于NC组织,E-cad蛋白表达水平低于NC组织,差异有统计学意义(P<0.05)。Caski+c-Src抑制剂组侵袭、迁移细胞计数及c-Src、FAK蛋白表达水平均低于Caski组,细胞凋亡率、E-cad蛋白表达水平高于Caski组,差异有统计学意义(P<0.05)。结论c-Src可能通过上调FAK和下调E-cad,导致宫颈癌细胞侵袭和迁移能力下降,其机制可能与凋亡增多有关。

Objective To study the role and mechanism of C-terminal Src protein(c-Src)/focal adhesion kinase(FAK)/E-cadherin(E-cad)pathway in invasion and metastasis of cervical cancer.Methods Thirty cervical squamous cell carcinoma(CSCC)tissue samples and 30 normal cervical(NC)tissue samples were collected from the First Affiliated Hospital of Hunan University of Medicine from January 2019 to December 2020.Immunohistochemical stain was used to detect the positive rate of E-cad in CSCC and NC tissue samples.The relationship between E-cad and clinicopathological features of CSCC patients was analyzed.Western blot was used to detect the protein expression levels of c-Src,FAK,and E-cad in CSCC and NC tissues.In addition,cervical cancer Caski cell lines were selected and divided into Caski group and Caski+c-SRC inhibitor group.The Caski group was not treated,and the Caski+c-SRC inhibitor group was added with c-Src inhibitor(2.7 nmol/L).The migration,invasion,and apoptosis of the two groups were observed,and the protein expression levels of c-Src,FAK,and E-cad in the two groups were detected by Western blot.Results The positive rate of E-cad in CSCC tissue samples was lower than that in NC,the difference was statistically significant(P<0.05).E-cad expression was correlated with International Federation of Gynecology and Obstetrics staging,differentiation degree,and recurrence of CSCC patients(P<0.05).The expression levels of c-Src and FAK protein in CSCC tissue samples were higher than those in NC tissue samples,while the expression level of E-cad protein was lower than that in NCT,the differences were statistically significant(P<0.05).The count of invaded and migrated cells and the expression levels of c-Src and FAK protein in Caski+c-Src inhibitor group were lower than those in Caski group,and the apoptosis rate and expres sion level of E-cad protein were higher than those in Caski group, the differences were statistically significant (P < 0.05). Conclusion C-src may lead to decreased invasion and migration ability of cervical cancer cells by up-regulating FAK and down-regulating E-cad, the mechanism may be related to increased apoptosis.