CAR-NK cell therapy inhibits the growth of gastric cancer xenografts with gastric cancer cell by regulating the PD-1/PD-L1 axis

ZHOU Jing-tao; LIU Jia; Nuermaimaiti·Amidula; LIU Yuan-yuan

doi:10.11659/jjssx.08E024109

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CAR-NK cell therapy inhibits the growth of gastric cancer xenografts with gastric cancer cell by regulating the PD-1/PD-L1 axis

Basic Research | 更新时间：2025-09-24

- CAR-NK cell therapy inhibits the growth of gastric cancer xenografts with gastric cancer cell by regulating the PD-1/PD-L1 axis
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 34, Issue 9, Pages: 747-752(2025)
- 作者机构：
  
  1.新疆医科大学第七附属医院普外科，新疆乌鲁木齐 830054
  2.新疆维吾尔自治区第八人民医院骨科，新疆乌鲁木齐 830000
  3.新疆医科大学第七附属医院药学部，新疆乌鲁木齐 830054
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.08E024109
  CLC： R735.2
- Received：27 August 2024，
  
  Published：25 September 2025
- 稿件说明：
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周京涛,刘佳,努尔买买提·阿米都拉等.CAR-NK细胞疗法通过调控PD-1/PD-L1轴抑制胃癌细胞异种移植瘤的生长[J].局解手术学杂志,2025,34(09):747-752.

ZHOU Jing-tao,LIU Jia,Nuermaimaiti·Amidula ,et al.CAR-NK cell therapy inhibits the growth of gastric cancer xenografts with gastric cancer cell by regulating the PD-1/PD-L1 axis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(09):747-752.
周京涛,刘佳,努尔买买提·阿米都拉等.CAR-NK细胞疗法通过调控PD-1/PD-L1轴抑制胃癌细胞异种移植瘤的生长[J].局解手术学杂志,2025,34(09):747-752. DOI： 10.11659/jjssx.08E024109.

ZHOU Jing-tao,LIU Jia,Nuermaimaiti·Amidula ,et al.CAR-NK cell therapy inhibits the growth of gastric cancer xenografts with gastric cancer cell by regulating the PD-1/PD-L1 axis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(09):747-752. DOI： 10.11659/jjssx.08E024109.

摘要

目的

探讨嵌合抗原受体（CAR）-自然杀伤（NK）细胞疗法对胃癌细胞和异种移植瘤生长的作用和潜在机制。

方法

细胞实验：将胃癌细胞系SGC7901和MGC803分别与CAR-NK细胞共培养作为CAR-NK组，将NK细胞分别与SGC7901和MGC803细胞共培养作为NK组，RT-qPCR检测2组细胞中PD-1和PD-L1的mRNA表达水平；用EdU染色法和CCK-8法检测细胞增殖能力；Transwell实验检测细胞迁移和侵袭能力，流式细胞术检测细胞周期变化，Western blot检测细胞中PD-1和PD-L1蛋白表达。动物实验：建立异种移植瘤小鼠模型，并分为空白对照组（接种常规培养的SGC7901细胞）、NK治疗组（联合接种SGC7901细胞和NK细胞）、CAR-NK治疗组（联合接种SGC7901细胞和CAR-NK细胞）、CAR-NK+rhPD-1治疗组（联合接种SGC7901细胞和CAR-NK细胞，同时腹腔注射rhPD-1 5 mg/kg）、CAR-NK+rhPD-L1治疗组（联合接种SGC7901细胞和CAR-NK细胞，同时腹腔注射rhPD-L1 5 mg/kg），每组5只。观察各组肿瘤体积并记录肿瘤质量，Western blot检测各组肿瘤组织中PD-1和PD-L1蛋白表达。

结果

与NK组比较，CAR-NK组SGC7901和MGC803细胞的增殖率、迁移和侵袭数均显著降低/减少（

0.05）。与NK组比较，CAR-NK组胃癌细胞S期数量增加（

0.05），

G2/M期数量减少（

0.05）。与NK组比较，CAR-NK组SGC7901细胞中PD-1和PD-L1的mRNA表达量和蛋白表达量均明显下调（

0.05）。在异种移植瘤小鼠模型中，与NK治疗组比较，CAR-NK治疗组肿瘤组织中PD-1和PD-L1的蛋白表达量下调，肿瘤体积减小，肿瘤质量减轻，差异均有统计学意义（

0.05）。与空白对照组比较，CAR-NK治疗组肿瘤组织中PD-1和PD-L1的蛋白表达量均下调，肿瘤体积减小，肿瘤质量减轻，差异均有统计学意义（

0.05）。与CAR-NK治疗组比较，CAR-NK+rhPD-1治疗组肿瘤组织PD-1蛋白表达上调，且肿瘤体积增大，肿瘤质量增加，差异均有统计学意义（

0.05）。与CAR-NK治疗组比较，CAR-NK+rhPD-L1治疗组肿瘤组织中PD-L1蛋白表达上调，且肿瘤体积增大，肿瘤质量增加，差异均有统计学意义（

0.05）。

结论

CAR-NK细胞疗法对胃癌细胞的增殖、迁移和侵袭有明显抑制作用，对胃癌细胞周期进程具有阻滞作用，其可能通过抑制PD-1/PD-L1轴抑制异种移植瘤的生长。

Abstract

Objective

To investigate the effect and potential mechanism of chimeric antigen receptor （CAR）-natural killer （NK） cell therapy on the growth of gastric cancer cells and xenograft tumors.

Methods

Cell experiments： The gastric cancer cell lines of SGC7901 and MGC803 were co-cultured with CAR-NK cells as the CAR-NK group，the NK cells were co-cultured with SGC7901 and MGC803 cells as the NK group，respectively. The mRNA levels of PD-1 and PD-L1 in both groups were detected by RT-qPCR. The cell proliferation ability was assessed using EdU staining and CCK-8 assay. The cell migration and invasion abilities were detected by Transwell assay. The change of cell cycle was detected by flow cytometry. The expression of PD-1 and PD-L1 proteins in cells was detected by Western blot. Animal experiments： Mice were established model of xenograft tumors and divided into the blank control group （inoculated with routinely cultured SGC7901 cells），NK treatment group （inoculated SGC7901 cells combined NK cells），CAR-NK treatment group （inoculated SGC7901 cells combined CAR-NK cells），CAR-NK+rhPD-1 treatment group （inoculated SGC7901 cells combined CAR-NK cells，with intraperitoneal injection of 5 mg/kg rhPD-1 concurrently），and CAR-NK+rhPD-L1 treatment group （inoculated SGC7901 cells combined CAR-NK cells，with intraperitoneal injection of 5 mg/kg rhPD-L1 concurrently），with 5 mice in each group. The tumor volume of each group was observed， the tumor weight was recorded，and the expression of PD-1 and PD-L1 proteins in the tumor tissue of each group were detected by Western blot.

Results

Compared to the NK group，the CAR-NK group showed significantly decreased proliferation rate，and numbers of migration and invasion of SGC7901 and MGC803 cells （

0.05）. Compared to the NK group，the number of S phase cells increased，while G2/M phase cells decreased in the CAR-NK group（

0.05）. Compared to the NK group，the mRNA and protein expression levels of PD-1 and PD-L1 significantly downregulated in SGC7901 cell of the CAR-NK group （

0.05）. In the xenograft mouse model，compared to the NK treatment group，the protein expression of PD-1 and PD-L1 downregulated in the tumor tissues of the CAR-NK treatment group，with smaller tumor volume and decreased tumor weight，the differences were statistically significant （

0.05）.Compared to the blank control group，the CAR-NK treatment group exhibited downregulated protein expression of PD-1 and PD-L1 in tumor tissues，reduced tumor volume，and decreased tumor weight，with statistically significant differences（

0.05）.Compared to the CAR-NK treatment group，the CAR-NK+rhPD-1 treatment group showed upregulated expression of PD-1 protein，large tumor volume，and increased tumor weight，with statistically significant differences （

0.05）. Compared to the CAR-NK treatment group，the CAR-NK+rhPD-L1 treatment group exhibited upregulated expression of PD-L1 protein，large tumor volume，and increased tumor weight，with statistically significant differences （

0.05）.

Conclusion

CAR-NK cell therapy have a significant inhibitory effect on the proliferation，migration，and invasion of gastric cancer cells，resulting in the gastric cancer cell cycle arrest，which may inhibit the growth of xenograft tumors by inhibiting the PD-1/PD-L1 axis.

关键词

Keywords

references

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