胎球蛋白A通过MAPK信号通路抑制卵巢癌迁移行为及机制研究

AHSG inhibits the migration of ovarian cancer through MAPK/ERK signaling pathway

目的探讨胎球蛋白A(AHSG)对卵巢癌细胞迁移行为的影响及分子机制。方法收取2022年9月至2023年1月于徐州市中心医院冻存的正常卵巢组织标本及卵巢癌组织标本。通过UALCAN数据库检索AHSG在卵巢癌中的表达情况;通过蛋白质印迹法检测临床卵巢癌组织及正常卵巢组织标本中AHSG蛋白表达水平,免疫组织化学染色检测卵巢癌及正常卵巢组织石蜡切片中AHSG蛋白表达水平;以卵巢癌细胞系SKOV3细胞、HO8910细胞作为研究对象,通过病毒转染技术构建上调AHSG表达的SKOV3细胞、HO8910细胞,通过伤口愈合实验、Transwell实验检测对照组及AHSG过表达组细胞迁移能力;蛋白质印迹法检测AHSG过表达后对促分裂原活化的蛋白质激酶(MAPK)通路关键蛋白c-Jun氨基端激酶(JNK)、磷酸化(p-)JNK、p38丝裂原激活的蛋白激酶(p38 MAPK)、p-p38 MAPK、胞外信号调节激酶(ERK)、p-ERK的影响。结果UALCAN数据库显示AHSG在卵巢癌中低表达;蛋白质印迹法结果显示卵巢癌组织中AHSG蛋白表达水平显著降低(P<0.05)。免疫组织化学染色实验显示AHSG蛋白表达水平在卵巢癌组织中显著下降(2.11±0.63比12.41±1.04,P<0.05)。伤口愈合实验显示过表达AHSG可显著抑制卵巢癌SKOV3细胞[(14.92±5.72)%比(45.07±7.07)%]、HO8910细胞[(56.31±2.37)%比(64.30±1.76)%]的迁移能力(P<0.05);Transwell实验显示过表达AHSG可显著抑制卵巢癌SKOV3细胞[(110.00±4.36)个比(201.33±23.18)个]、HO8910细胞[(119.00±16.64)个比(167.00±6.08)个]的迁移能力(P<0.05)。上调AHSG可显著抑制MAPK信号通路关键蛋白的表达,在SKOV3细胞中,p-p38 MAPK/p38 MAPK(1.03±0.22比1.67±0.15)、p-JNK/JNK(0.75±0.27比1.25±0.15)、p-ERK/ERK(0.55±0.11比1.01±0.22)(P<0.05)。同样在HO8910细胞中,p-p38 MAPK/p38 MAPK(0.42±0.02比1.11±0.28)、p-JNK/JNK(0.73±0.12比0.99±0.11)、p-ERK/ERK(0.94±0.20比1.69±0.13)(P<0.05)。结论AHSG可以抑制卵巢癌细胞迁移能力,其作用机制可能与AHSG抑制MAPK通路的激活有关。

Objective To investigate the effect of Alpha-2 Heremans Schmid Glycoprotein(AHSG)on the migration of ovarian cancer cells and its molecular mechanism.Methods The collection of normal ovarian tissue specimens and ovarian cancer tissue specimens were cryopreserved at Xuzhou Central Hospital from September 2022 to January 2023.The expression of AHSG in ovarian cancer was retrieved from the UALCAN database;The expression levels of AHSG in clinical ovarian cancer tissues and normal ovarian tissues were detected by Western blot experiment.And the expression of AHSG protein in paraffin sections of ovarian cancer and normal ovarian tissues were detected by the immunohistochemical method.SKOV3 cells and HO8910 cells were selected as the research subjects.SKOV3 cells and HO8910 cells with up-regulated AHSG expression were constructed by virus transfection technique.The migration ability of control group and AHSG overexpression group was detected by wound healing assay and Transwell assay.Western blotting analysis was used to detect the effects of overexpression of AHSG on the MAPK pathway key proteins,Jun N-terminal kinase(JNK),phosphorylated Jun N-terminal kinase(p-JNK),p38 mitogen-activated protein kinase(p38 MAPK),phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK),extracellular signal-regulated kinase(ERK),and phosphorylated extracellular signal-regulated kinase(p-ERK).Results UALCAN database showed the low expression of AHSG in ovarian cancer.Compared with normal ovarian tissues,Western blot showed that the expression level of AHSG protein was significantly decreased in ovarian cancer tissues(P<0.05).Immunohistochemical method showed that the expression of AHSG protein in the ovarian cancer group was lower than the normal group(2.11±0.63 vs.12.41±1.04,P<0.05).The wound healing experiment showed that overexpression of AHSG could significantly inhibit the migration ability of SKOV3[(14.92±5.72)%vs.(45.07±7.07)%]and HO8910 cells[(56.31±2.37)%vs.(64.30±1.76)%](P<0.05).Transwell assay also showed that overexpression of AHSG could significantly inhibit the migration ability of SKOV3(110.00±4.36 vs.201.33±23.18)and HO8910 cells(119.00±16.64 vs.167.00±6.08)(P<0.05).Up-regulation of AHSG could significantly inhibit the key proteins of MAPK signaling pathway.The relative expression of p-p38 MAPK/p38 MAPK(1.03±0.22 vs.1.67±0.15),p-JNK/JNK(0.75±0.27 vs.1.25±0.15),p-ERK/ERK(0.55±0.11 vs.1.01±0.22)in the SKOV3 cells(P<0.05).Similarly,the relative expression of p-p38 MAPK/p38 MAPK(0.42±0.02 vs.1.11±0.28),p-JNK/JNK(0.73±0.12 vs.0.99±0.11),p-ERK/ERK(0.94±0.20 vs.1.69±0.13)were also found in the HO8910 cells(P<0.05).Conclusion AHSG can inhibit ovarian cancer cell migration,and its mechanism may be related to AHSG can inhibit the activation of MAPK pathway.