摘要
目的探讨去泛素化酶含约瑟芬结构域2蛋白(JOSD2)在乳腺癌组织中表达情况及对术后中医治疗敏感性和乳腺癌细胞增殖的影响。方法①选择2021年1月—2022年1月在中国中医科学院广安门医院南区行乳腺癌手术后患者100例作为乳腺癌组,选取同期年龄与乳腺癌组患者匹配的乳腺良性肿瘤患者80例作为乳腺良性肿瘤组,酶联免疫吸附实验检测2组血清JOSD2水平,免疫组化检测乳腺癌组织、癌旁组织、乳腺良性病变组织中JOSD2阳性表达情况;根据JOSD2表达情况将乳腺癌患者分为JOSD2阳性组和JOSD2阴性组,比较2组中医治疗前后中医证候积分、免疫功能指标(CD3^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))、外周血象(白细胞计数、血小板计数、血红蛋白)、血清肿瘤标记物[癌胚抗原(CEA)、糖蛋白抗原153(CA153)]及细胞因子[血管内皮生长因子(VEGF)、p53基因(p53)]水平。②取乳腺癌细胞系HCC1937和人永生化正常乳腺上皮细胞MCF-10A,实验设MCF-10A组、HCC1937组、sh-NC-HCC1937组和sh-JOSD2-HC C1937组,每组细胞相应培养处理后,采用Western blot法检测细胞中JOSD2蛋白表达情况;实验设HCC1937组、sh-NC-HCC1937组和sh-JOSD2-HCC1937组,CCK-8实验和EdU实验检测HCC1937细胞增殖情况,碘化丙啶染色法检测细胞周期。结果①乳腺癌组患者治疗前后血清JOSD2水平均明显高于乳腺良性肿瘤组(P均<0.05),JOSD2在乳腺癌组织中染色较癌旁组织和乳腺良性肿瘤组织明显偏深;治疗后乳腺癌组患者血清JOSD2水平明显低于治疗前(P<0.05)。JOSD2阳性组和JOSD2阴性组患者中医治疗后中医证候积分及血清CA153、CEA、VEGF、p53水平均明显低于治疗前(P均<0.05),但JOSD2阳性组患者上述指标均明显高于同期JOSD2阴性组(P均<0.05);JOSD2阳性组和JOSD2阴性组患者中医治疗后CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)及白细胞计数、血小板计数、血红蛋白水平均明显高于治疗前(P均<0.05),但JOSD2阳性组患者上述指标均明显低于同期JOSD2阴性组(P均<0.05)。②HCC1937组JOSD2蛋白相对表达量明显高于MCF-10A组(P<0.05),sh-JOSD2-HCC1937组均明显低于HCC1937组与sh-NC-HCC1937组(P均<0.05)。与HCC1937组和sh-NC-HCC1937组比较,sh-JOSD2-HCC1937组细胞光密度值和细胞增殖活性均明显降低(P均<0.05),细胞G_(0)/G_(1)期细胞比例明显增高(P均<0.05),S期和G_(2)/M期细胞比例明显降低(P均<0.05)。结论乳腺癌患者血清和乳腺癌组织中JOSD2表达显著上调,JOSD2表达对术后中医治疗敏感,且沉默JOSD2对乳腺癌细胞增殖具有抑制作用,JOSD2有望成为乳腺癌早期筛查、诊断、治疗的新型生物标志物与靶点。
Objective It is to investigate the expression of deubiquitinating enzyme Josephin domain-containing protein 2(JOSD2)in breast cancer tissue and its effect on the postoperative treatment with traditional Chinese medicine and proliferation of breast cancer cells.Methods①100 patients after breast cancer surgery in the South Branch of Guang'anmen Hospital of China Academy of Traditional Chinese Medicine from January 2021 to January 2022 were selected as the breast cancer group,and 80 patients with benign breast tumors whose age matched that of patients in the breast cancer group during the same period were selected as the benign breast tumor group.The level of serum JOSD2 in patients of the two groups was detected by enzyme-linked immunosorbent assay,and the positive expressions of JOSD2 in breast cancer tissues,paracancerous tissues,and benign breast lesion tissues were detected by immunohistochemistry.According to JOSD2 expression,the breast cancer patients were divided into JOSD2-positive and JOSD2-negative groups,and the scores of traditional Chinese medicine syndromes,the levels of immune function indexes(CD3^(+),CD4^(+),CD8^(+),and CD4^(+)/CD8^(+)),and peripheral blood examination(leukocyte count,platelet count,hemoglobin),serum tumor markers[carcinoembryonic antigen(CEA),glycoprotein antigen 153(CA153)]and cytokines[vascular endothelial growth factor(VEGF),p53 gene(p53)]were compared before and after between the two groups.②Breast cancer cell line HCC1937 and human immortalized normal mammary epithelial cells MCF-10A were taken,MCF-10A group,HCC1937 group,sh-NC-HCC1937 group and sh-JOSD2-HCC1937 group were set up in the experiments,the cells in each group were cultured accordingly,and then the protein expression of JOSD2 in the cells were detected by Western blot;HCC1937,sh-NC-HCC1937 and sh-JOSD2-HCC1937 groups were set up in the experiments,the proliferation of HCC1937 cells was detected by CCK-8 assay and EdU assay,and the cell cycle was detected by propidium iodide staining.Results①The serum level of JOSD2 in patients of the breast cancer group before and after treatment were significantly higher than that in patients of the benign breast tumor group(P<0.05),and the staining of JOSD2 in breast cancer tissues was significantly darker than that in paracarcinoma tissues and benign breast tumor tissues;the serum level of JOSD2 in patients of the breast cancer group were significantly lower than that before treatment(P<0.05).The scores of traditional Chinese medicine syndromes and serum levels of CA153,CEA,VEGF and p53 in patients of the JOSD2-positive and JOSD2-negative groups were significantly lower after treatment with traditional Chinese medicine than those before treatment(all P<0.05),but the above indicators in patients of the JOSD2-positive group were significantly higher than those of the JOSD2-negative group in the same period(all P<0.05);the levels of CD3^(+),CD4^(+),CD4^(+)/CD8^(+)and the levels of white blood cell count,platelet count and hemoglobin in patients of the JOSD2-positive and JOSD2-negative groups were significantly higher after treatment with traditional Chinese medicine than those before treatment(all P<0.05),but the above indexes in patients of the JOSD2-positive group were significantly lower than those of the JOSD2-negative group in the same period(all P<0.05).②The relative protein expression of JOSD2 in the HCC1937 group was significantly higher than that in the MCF-10A group(P<0.05),and the expression in the sh-JOSD2-HCC1937 group was significantly lower than that in the HCC1937 group and sh-NC-HCC1937 group(both P<0.05).Compared with both HCC1937 group and sh-NC-HCC1937 group,the cell optical density value and cell proliferation activity of sh-JOSD2-HCC1937 group were significantly lower(both P<0.05),and the proportion of cells in G_(0)/G_(1)phase was significantly higher(both P<0.05),and the proportion of cells in S phase and G_(2)/M phase was significantly lower(both P<0.05).Conclusion JOSD2 expression is significantly up-regulated in serum and breast cancer tissues of breast cancer patients,and it is sensitive to postoperative treatment with traditional Chinese medicine,silencing JOSD2 can inhibit proliferation of breast cancer cells,and JOSD2 is expected to become a novel biomarker and target for early screening,diagnosis,and treatment of breast cancer.
作者
许华颖
王如然
张玉杰
李宏燕
XU Huaying;WANG Ruran;ZHANG Yujie;LI Hongyan(South Branch of Guang’anmen Hospital of China Academy of Chinese Medical Sciences,Beijing 102600,China)
出处
《现代中西医结合杂志》
2025年第2期147-154,共8页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
北京中医药科技发展基金项目(JJ2018-59)。
关键词
乳腺癌
去泛素化酶含约瑟芬结构域2蛋白
细胞增殖
中医
breast cancer
deubiquitinating enzyme Josephin domain-containing protein 2
cell proliferation
Chinese medicine
