基膜聚糖通过抑制1型辅助T细胞分化提高耐药性卵巢癌治疗效果

Lumican enhanced the therapeutic effect of cisplatin-resistant ovarian cancer by inhibiting the differentiation of Th1 cells

目的探讨基膜聚糖(LUM)在卵巢癌顺铂耐药中的作用机制,并初步探索其对1型辅助T(Th1)细胞分化的影响。方法通过基因表达数据集(GEO)筛选卵巢癌顺铂耐药株及敏感株中差异表达的基因,通过实时定量PCR检测其表达水平。利用小干扰RNA(siRNA)敲低人卵巢癌细胞中LUM表达,并用Western blot法验证敲低效率。通过流式细胞术检测敲低LUM对顺铂处理后的卵巢癌细胞系周期及凋亡的影响。通过蛋白质相互作用(PPI)网络筛选LUM潜在靶蛋白,并用分子对接验证其相互作用。肿瘤免疫评估资源数据库(TIMER)筛选LUM对卵巢癌系分泌细胞因子的影响,并利用ELISA与实时定量PCR验证。流式细胞术分析LUM对共培养的CD4^(+)T细胞分化的调控作用。结果GEO数据显示LUM在顺铂耐药株中显著上调,且其表达水平与患者预后相关。LUM在卵巢癌顺铂耐药细胞株中表达水平较高,且顺铂治疗能促进LUM的表达。敲低LUM增加顺铂诱导的卵巢癌细胞的凋亡和细胞周期阻滞,提高药物敏感性。靶基因筛选表明,LUM可能通过与Src同源域磷酸酶2(SHP2)相互作用调控卵巢癌细胞对顺铂的敏感性。另外,TIMER分析结果提示高表达LUM抑制Th1细胞分化,在卵巢癌耐药株中敲低LUM促进Th1细胞分化,通过调控γ干扰素(IFN-γ)和白细胞介素12(IL-12)分泌影响免疫细胞的肿瘤杀伤力。结论LUM在卵巢癌顺铂耐药中表达上调,可能通过调控SHP2相关信号通路降低卵巢癌细胞顺铂敏感性。LUM还通过调控卵巢癌细胞细胞因子分泌,进而抑制Th1细胞分化,促进肿瘤耐药性产生,可能是卵巢癌治疗新的潜在靶点。

Objective To investigate the mechanism of the basement membrane proteoglycan lumican(LUM)in cisplatin resistance in ovarian cancer and to preliminarily explore its effect on type 1 T helper(Th1)cell differentiation.Methods Differentially expressed genes(DEGs)between cisplatin-resistant and cisplatin-sensitive ovarian cancer cell lines were screened using the public gene expression database(GEO).The expression levels of these genes were detected by RT-qPCR.LUM expression in human ovarian cancer cells was knocked down using small interfering RNA(siRNA),and the knockdown efficiency was verified by Western blotting.Flow cytometry was used to detect the effects of LUM knockdown on the cell cycle and apoptosis of cisplatin-treated ovarian cancer cell lines.Potential target proteins of LUM were screened through the PPI network,and their interactions were validated by molecular docking.The TIMER database was used to screen the effects of LUM on cytokine secretion in ovarian cancer cell lines,and the results were validated by ELISA and RT-qPCR.Flow cytometry was performed to analyze the regulatory effect of LUM on the differentiation of CD4^(+)T cells.Results GEO data showed that LUM was significantly upregulated in cisplatin-resistant cell lines,and its expression level was correlated with patient prognosis.LUM expression level was higher in that of cisplatin-resistant ovarian cancer cell lines,and cisplatin treatment promoted LUM expression.Knockdown of LUM increased cisplatin-induced apoptosis and cell cycle arrest in ovarian cancer cells,enhancing drug sensitivity.Target gene screening suggested that LUM might regulate cisplatin sensitivity in ovarian cancer cells through interaction with Src homology region phosphatase 2(SHP2).Additionally,TIMER database analysis suggested that high LUM expression inhibited Th1 cell differentiation.Knockdown of LUM in cisplatin-resistant cell lines promoted Th1 cell differentiation by regulating the secretion of interferonγ(IFN-γ)and interleukin 12(IL-12)cytokines,thereby influencing the tumoricidal activity of immune cells.Conclusion LUM is upregulated in cisplatin-resistant ovarian cancer cells and reduces cisplatin sensitivity in ovarian cancer cells by regulating the SHP2-related signaling pathway.LUM also promotes tumor resistance by inhibiting Th1 cell differentiation through the regulation of cytokine secretion by ovarian cancer cells,making it a potential target for ovarian cancer treatment.