华蟾素胶囊联合美施康定对肺腺癌EGFR突变骨转移癌痛患者中circRNAs差异表达的 影响

Effects of Huachansu Capsules Combined with MS Contin on Differentially Expressed circRNAs in Plasma of Patients with EGFR-Mutant Lung Adenocarcinoma and Bone Metastasis-Related Cancer Pain

摘要: 目的：筛选并分析华蟾素胶囊联合美施康定治疗肺腺癌EGFR突变骨转移癌痛患者血浆中差异表达的环状RNA (circular RNA, circRNAs)，通过临床数据追踪、生物信息学方法探讨该联合方案调控癌痛的潜在分子机制。方法：选取肺腺癌EGFR突变骨转移癌痛患者60例，随机分为联合治疗组和单一用药组，每组各30例；另从两组中各随机选取6例患者作为测序亚组，采集测序亚组患者治疗前后血浆标本，用于circRNAs高通量测序分析。追踪两组患者疼痛达标时间、美施康定日均用量、不良反应发生率。采用高通量测序技术检测测序亚组血浆circRNAs表达谱，以|log2(FC)| > 1.5且FDR < 0.05为标准筛选差异表达circRNAs，组间比较采用t检验，并对差异circRNAs进行生物信息学分析。分析临床疗效指标与关键circRNAs表达变化的相关性。结果：联合治疗组疼痛达标时间显著短于单一用药组，美施康定日均用量显著低于单一用药组，不良反应总发生率显著低于单一用药组(P < 0.05)。联合治疗组与单一用药组相比，共筛选出710个差异表达circRNAs (P < 0.05)，其中326个上调，384个下调。GO分析显示，差异circRNAs亲本基因主要富集于转录调控活性、蛋白质翻译后修饰、细胞内信号转导调控等生物学过程与分子功能；KEGG通路分析提示，差异circRNAs主要富集于癌症相关通路、激素信号转导、物质代谢与转运等通路。结论：华蟾素胶囊联合美施康定可显著提升镇痛效率、减少阿片类药物用量并降低不良反应，并可显著改变肺腺癌EGFR突变骨转移癌痛患者血浆circRNAs表达谱。circRNAs可能作为联合治疗调控网络中的关键节点，通过重塑miRNA功能取向，介导中药与阿片类药物协同抗肿瘤及镇痛效应。hsa_circ_0039161、hsa_circ_0001707等关键circRNAs有望成为疗效评估潜在生物标志物，为阐明中西医结合治疗癌痛的分子机制及实现癌痛精准治疗提供新的理论依据与研究方向。

Abstract: Objective: To screen and analyze the differentially expressed circular RNAs (circRNAs) in the plasma of patients with EGFR-mutant lung adenocarcinoma complicated with bone metastasis-related cancer pain who were treated with Huachansu Capsules combined with MS Contin, and to explore the potential molecular mechanism of this combined regimen in regulating cancer pain through clinical data tracking and bioinformatics methods. Methods: A total of 60 patients with EGFR-mutant lung adenocarcinoma and bone metastasis-related cancer pain were enrolled and randomly divided into a combined treatment group and a monotherapy group, with 30 cases in each group. Additionally, 6 patients were randomly selected from each group to form a sequencing subgroup, and plasma samples of patients in the sequencing subgroup were collected before and after treatment for high-throughput sequencing analysis of circRNAs. The pain relief time, daily dosage of MS Contin and incidence of adverse reactions were tracked in both groups. High-throughput sequencing technology was used to detect the plasma circRNA expression profiles of the sequencing subgroup. Differentially expressed circRNAs were screened with the criteria of |log2(FC)| > 1.5 and FDR < 0.05, independent samples t-test was used for inter-group comparison, and bioinformatics analysis was performed on the differentially expressed circRNAs. The correlation between clinical efficacy indicators and the expression changes of key circRNAs was analyzed. Results: The pain relief time of the combined treatment group was significantly shorter than that of the monotherapy group, the total consumption of MS Contin equivalent dose was significantly lower than that of the monotherapy group, and the total incidence of adverse reactions was significantly lower than that of the monotherapy group (P < 0.05). A total of 710 differentially expressed circRNAs were identified in the combined treatment group compared with the monotherapy group (P < 0.05), including 326 up-regulated and 384 down-regulated ones. Gene Ontology (GO) analysis showed that the parental genes of the differentially expressed circRNAs were mainly enriched in biological processes and molecular functions such as transcriptional regulatory activity, post-translational protein modification, and regulation of intracellular signal transduction. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that these differentially expressed circRNAs were mainly enriched in cancer-related pathways, hormone signal transduction, substance metabolism and transport pathways, etc. Conclusion: Huachansu Capsules combined with MS Contin can significantly improve analgesic efficiency, reduce opioid dosage and decrease adverse reactions, and also remarkably alter the plasma circRNA expression profile in patients with EGFR-mutant lung adenocarcinoma and bone metastasis-related cancer pain. circRNAs may serve as key nodes in the regulatory network of the combined therapy, and mediate the synergistic anti-tumor and analgesic effects of traditional Chinese medicine combined with opioids by reshaping the functional orientation of miRNAs. Key circRNAs such as hsa_circ_0039161 and hsa_circ_0001707 are expected to be potential biomarkers for therapeutic efficacy evaluation, which provides a new theoretical basis and research direction for clarifying the molecular mechanism of integrated traditional Chinese and Western medicine in the treatment of cancer pain and realizing precise treatment of cancer pain.