目的：明确康莱特注射液（KLTi）通过调控胆固醇代谢对肺腺癌A549 细胞恶性生物学行为的抑制作用。方法：构建A549 细胞体外培养模型，设置空白对照组（CON组）、KLTi组、顺铂（DDP）组及KLTi+DDP 组，分别给予对应药物干预，采用CCK-8法检测不同分组的药物干预对A549 细胞增殖的影响，并确定IC50 值用于后续实验；使用细胞划痕实验、平板克隆形成实验、 Transwell 侵袭实验观察不同分组药物对A549 细胞恶性生物学行为的影响；流式细胞术检测不同分组药物对A549 细胞晚期凋亡水平的影响；WB法检测各组细胞上皮间质转化（EMT）相关蛋白表达，ELISA 法检测促炎因子释放水平。采用比色法检测细胞胆固醇含量水平的组间差异，借助WB法检测胆固醇代谢相关膜通道蛋白ATP结合盒转运蛋白A1（ABCA1）及功能蛋白ATP柠檬酸裂解酶（ACLY）、肽基脯氨酰异构酶B（PPIB）表达水平差异。结果：KLTi 及DDP对A549 细胞抑制作用具有时间及剂量依赖性（均P<0.05），最终选择2 mg/mL KLTi、3 μg/mL DDP作为干预剂量，按分组加药干预48 h后显示，KLTi 单用或联合DDP均可抑制A549 细胞克隆形成、迁移、侵袭能力且促进其晚期凋亡，KLTi+DDP 组的效果更加明显（P<0.05 或P<0.01）； KLTi 单用或联合DDP 可通过调控E-cadherin、vimentin、snail 蛋白表达从而影响A549 细胞EMT 进程（P<0.05 或P<0.01），同时下调IL-6 及IL-8的释放水平（P<0.05 或P<0.01）。KLTi 单用及联合DDP 均可以明显降低A549 细胞胆固醇含量（P<0.05 或P<0.01），并且对ABCA1、ACLY、PPIB 表达具有调控作用，联合组的效果更加明显（P<0.05 或P<0.01）。结论：KLTi 可能通过调控胆固醇代谢水平及相关通道蛋白抑制肺腺癌A549细胞增殖、迁移、侵袭等恶性生物学行为及EMT进程。

Objective： To investigate the inhibitory effect of Kanglaite injection (KLTi) on the biological behaviors of lung adenocarcinoma A549 cells by regulating cholesterol metabolism. Methods: An in vitro monoculture model of A549 cells was established. A blank control group (CON group), a KLTi group, a cisplatin group (DDP group) and a KLTi + DDP group were set and given corresponding drug intervention, respectively. CCK-8 method was used to detect the effects of different interventions on the proliferation of A549 cells, and IC50 values were determined for subsequent experiments. The effects of different drugs on the malignant biological behaviors of A549 cells were compared by cell scratch assay, plate clonogenesis assay and Transwell invasion assay, and the late apoptosis of A549 cells were detected by flow cytometry. The expression of epithelial-mesenchymal transition (EMT) related proteins was detected by WB method, and the release level of pro-inflammatory factors was detected by ELISA. Colorimetric method was used to detect the difference of cholesterol content in 106 cells among groups. The difference in the expression levels of membrane channel protein ATP binding cassette transport protein A1 (ABCA1) and functional proteins ATP citrate lyase (ACLY) and peptidylprolyl isomerase B (PPIB) related to cholesterol metabolism was determined by using WB assay. Results: KLTi and DDP inhibited A549 cells in a time- and dose-dependent manner (both P<0.05). Finally, 2 mg/mL KLTi, 3 μg/mL DDP and 48 h of intervention were chosen as intervention dose for follow-up experiments. KLTi alone or combined with DDP could inhibit the clone formation, migration and invasion and promote the late apoptosis of A549 cells, with more prominent effect in the KLTi+DDP group (P<0.05 or P<0.01). KLTi alone or in combination with DDP could improve the EMT process of A549 cells by regulating the proteinexpression of E-cadherin, vimentin and snail (P<0.05 or P<0.01) and down-regulate the release of pro-inflammatory cytokines IL-6 and IL-8 (P<0.05 or P<0.01). KLTi alone or in combination with DDP could significantly reduce the cholesterol content of A549 cells (P<0.05 or P<0.01) and had a regulatory effect on ABCA1, ACLY and PPIB, with more prominent effects in the KLTi+DDP group (P<0.05 or P<0.01). Conclusion: KLTi inhibits the proliferation, migration, invasion and EMT process of lung adenocarcinoma A549 cells possibly by regulating cholesterol metabolism levels and related channel proteins.

国家重点研发计划-中医药现代化研究专项（No. 2018YFC1707405）；中央高水平中医医院临床研究和成果转化能力提升项目-临床科研一体化人才专项（创新团队培育项目）（No.HLCMHPP2023001)