中药单体川楝素介导hsa-circ-0025583/hsa-miR-671-5p/SYNGR2轴调控乳腺癌生长的分子机制研究

Molecular Mechanism of Chinese Traditional Medicine Monomer Toosendanin Mediating Growth of Breast Cancer by hsa-circ-0025583/hsa-miR-671-5p/SYNGR2 Axis

目的 探究中药单体川楝素介导的hsa-circ-0025583/hsa-miR-671-5p/SYNGR2轴调控乳腺癌生长的分子机制。方法 选取南通第三人民医院乳腺外科于2020年4月—2022年4月收治的乳腺癌患者40例为研究对象,患者均在医院进行手术,取患者乳腺癌及癌旁组织作为实验材料,采用定量荧光聚合酶链反应(quantitative polymerase chain reaction, qPCR)对其hsa-circ-0025583及SYNGR2的表达水平进行测定;取乳腺癌细胞株T47D细胞,加入不同浓度的川楝素,比较不同浓度川楝素对细胞增殖的抑制率;对所选乳腺癌细胞株系进行预处理,A组不进行特殊处理,B组中加入0.5μmol/L的川楝素,C组对乳腺癌细胞进行转染,敲除hsa-miR-671-5p,并向其中加入0.5μmol/L的川楝素,比较其hsa-circ-0025583及SYNGR2的表达水平,上述实验重复3次。结果 与正常癌旁组织相比较,乳腺癌组织中提取的hsa-circ-0025583及SYNGR2 mRNA表达水平均降低,差异有统计学意义(P<0.05)。hsa-circ-0025583及SYNGR2 mRNA在乳腺癌组织中均低表达,具有正相关性(r=0.965,P<0.05)。随着川楝素浓度的升高,乳腺癌细胞增殖的抑制率逐渐升高,且与0μmol/L浓度组比较均具有统计学意义(P<0.05)。对数据进行处理,得出乳腺癌细胞在24 h及48 h的半数抑制浓度分别为0.78μmol/L、0.49μmol/L。与A、C组相比较,B组的hsa-circ-0025583及SYNGR2 mRNA的表达水平明显升高,差异有统计学意义(P<0.05);但A、C组间比较,差异无统计学意义(P>0.05)。与A、C组相比较,B组24 h及48 h侵袭细胞数量明显降低,差异有统计学意义(P<0.05);但A、C组间侵袭细胞数量比较,差异无统计学意义(P>0.05)。结论 川楝素通过介导hsa-circ-0025583/hsa-miR-671-5p/SYNGR2轴调控hsa-circ-0025583及SYNGR2水平,进而影响乳腺癌细胞的增殖及侵袭能力,抑制乳腺癌细胞的生长。

Objective To explore the molecular mechanism of hsa-circ-0025583/hsa-miR-671-5p/SYNGR2 axis regu-lating the growth of breast cancer mediated by toosendanin.Methods Forty patients with breast cancer admitted to the Breast and Thyroid Surgery Department of Nantong Third People's Hospital from April 2020 to April 2022 were selected as the study sub-jects.All patients underwent surgery in the hospital.The breast cancer and adjacent tissues of patients were taken out as the ex-perimental materials.The expression levels of hsa-circ-0025583 and SYNGR2 were determined by quantitative fluorescent pol-ymerase chain reaction(qPCR).Breast cancer cell line T47D cells were added with different concentrations of toosendanin to compare the inhibition rate of toosendanin at different concentrations on cell proliferation.The selected breast cancer cell lines were pretreated.Group A did not receive special treatment.0.5μmol/L toosendanin was added to group B and group C trans-fected breast cancer cells,knocking out hsa-miR-671-5p and adding 0.5μmol/L toosendanin to them.The hsa-circ-0025583 and SYNGR2 expression levels were compared.The above experiments were repeated three times.Results Compared with those in the normal adjacent tissues,the expression levels of hsa-circ-0025583 and SYNGR2 mRNA were decreased,and the difference was significant(P<0.05).Both hsa-circ-0025583 and SYNGR2 mRNA were poorly expressed in breast cancer tissues,with a positive correlation(r=0.965,P<0.05).With the increase of tranazmabelin concentration,the inhibition rate of breast cancer cell proliferation gradually increased,and it was statistically different from the 0μmol/L concentration group(P<0.05).The data was processed such that the half-inhibitory concentrations of breast cancer cells were 0.78μmol/L and 0.49μmol/L at 24 h and 48 h,respectively.Compared with those of group A and C,the expression levels of hsa-circ-0025583 and SYNGR2 mRNA were significantly increased,and the difference was significant(P<0.05).However,there was no significance between group A and C(P>0.05).Compared with that of group A and C,the number of invaded cells at 24 h and 48 h was sta-tistically lower(P<0.05).However,the number of invasive cells between group A and C had no significance(P>0.05).Con-clusion Toosendanin regulates the levels of hsa-circ-0025583 and SYNGR2 by mediating the hsa-circ-0025583/hsa-miR-671-5p/SYNGR2 axis,thereby affecting the proliferation and invasion of breast cancer cells and inhibiting the growth of breast cancer cells.