SPART通过脂噬对胃癌细胞增殖和迁移能力的影响

doi:10.3760/cma.j.cn371439-20250612-00011

摘要/Abstract

摘要：

目的探讨SPART在胃癌组织中的表达情况及其对胃癌细胞增殖、迁移能力及脂噬的影响。方法通过生物信息学分析TCGA数据库中胃癌转录组数据，分析SPART的表达情况；选取2024年4月至2025年4月在新疆医科大学附属肿瘤医院确诊为胃癌并行手术治疗的5例患者，采用蛋白质印迹法检测其胃癌及癌旁组织中SPART蛋白表达水平。人胃癌细胞株AGS、NUGC3实验组均转染SPART过表达质粒，对照组均转染空白载体，采用蛋白质印迹法检测过表达SPART对微管相关蛋白轻链3（LC3）、溶酶体相关膜蛋白2（LAMP2）、围脂滴蛋白3（PLIN3）表达的影响。采用CCK-8、Transwell实验分别检测细胞增殖及迁移能力，免疫荧光实验检测SPART过表达后脂滴与LC3的含量。结果生物信息学分析提示，SPART在375例胃癌组织和32例正常胃黏膜组织中的表达量分别为3.06（1.62）、3.54（2.68），差异有统计学意义（U=7 429.00，P=0.025）。27对配对样本中，SPART在胃癌组织中的相对表达量为2.78±0.82，低于正常胃组织的3.84±1.22，差异有统计学意义（t=-3.80，P<0.001）。人类蛋白图谱数据库中的免疫组织化学染色结果显示，23例胃癌组织样本中，16例为SPART低表达（69.57%），7例中表达（30.43%），无高表达（0）。蛋白质印迹法结果显示，临床样本中胃癌组织SPART蛋白相对表达量为0.56（0.68），低于癌旁组织的1.38（1.05），差异有统计学意义（U=4.00， P=0.016）。CCK-8实验结果显示，第48小时，AGS细胞对照组、实验组的吸光度（A）值分别为0.52±0.01、0.41±0.01；NUGC3细胞对照组、实验组的A值分别为0.67±0.02、0.51±0.01，差异均有统计学意义（t=13.47， P<0.001；t=12.39， P<0.001）；第72小时，AGS细胞对照组、实验组的A值分别为0.82±0.01、0.67±0.01；NUGC3细胞对照组、实验组的A值分别为1.05±0.01、0.78±0.02，差异均有统计学意义（t=18.37， P<0.001；t=20.91， P<0.001）；第96小时，AGS细胞对照组、实验组的A值分别为1.04±0.03、0.74±0.01；NUGC3细胞对照组、实验组的A值分别为1.80±0.02、1.41±0.08，差异均有统计学意义（t=16.43， P<0.001；t=8.19， P=0.004）。Transwell实验结果显示，AGS、NUGC3细胞实验组中迁移细胞数分别为（500±17）、（276±21）个，均少于对照组的（771±33）、（327±12）个，差异均有统计学意义（t=-12.65，P<0.001；t=-3.65，P=0.030）。蛋白质印迹法结果显示，AGS细胞对照组、实验组LC3蛋白表达水平分别为0.47±0.08、1.06±0.32，LAMP2蛋白表达水平分别为0.65±0.03、0.98±0.11，PLIN3蛋白表达水平分别为0.82±0.15、0.43±0.10，差异均有统计学意义（t=3.11， P=0.040；t=5.02， P=0.007；t=3.75， P=0.020）。NUGC3细胞对照组、实验组LC3蛋白表达水平分别为0.89±0.04、1.45±0.16，LAMP2蛋白表达水平分别为0.68±0.18、1.22±0.22，PLIN3蛋白表达水平分别为1.71±0.45、0.35±0.06，差异均有统计学意义（t=5.89， P=0.004；t=3.29， P=0.030；t=5.19， P=0.007）。免疫荧光实验结果显示，AGS细胞对照组与实验组脂滴水平分别为1.08±0.09、0.32±0.03，LC3水平分别为0.40±0.01，0.88±0.15，差异均有统计学意义（t=13.88，P<0.001；t=-5.53，P=0.033）。结论 SPART在胃癌中低表达，过表达SPART可抑制胃癌细胞增殖和迁移并激活细胞脂噬。

关键词: 胃肿瘤, 细胞增殖, 细胞运动, 脂噬, SPART

Abstract:

Objective To investigate the expression of SPART in gastric cancer tissues and its effects on lipophagy， as well as proliferation and migration in gastric cancer cells. Methods Bioinformatics analysis was performed on gastric cancer transcriptome data from the TCGA database to assess SPART expression level. Five patients diagnosed with gastric cancer and undergoing surgical treatment at the Affiliated Tumor Hospital of Xinjiang Medical University from April 2024 to April 2025 were selected， and Western blotting was used to measure SPART protein levels in gastric cancer and adjacent normal tissues. The experimental groups of human gastric cancer cell lines AGS and NUGC3 were all transfected with SPART overexpression plasmids， while the control groups were all transfected with blank vectors. Western blotting was used to detect the effects of overexpressing SPART on the expression of microtubule-associated protein light chain 3 （LC3）， lysosome-associated membrane protein 2 （LAMP2）， and perilipin 3 （PLIN3）. Cell proliferation and migration capabilities were assessed using CCK-8 and Transwell assays， respectively. Immunofluorescence was used to examine lipid droplet content and LC3 levels after SPART overexpression. Results Bioinformatics analysis revealed that the expression levels of SPART in 375 gastric cancer tissues and 32 normal gastric mucosa tissues were 3.06 （1.62） and 3.54 （2.68）， respectively， with a statistically significant difference （U=7 429.00， P=0.025）. Among the 27 pairs of paired samples， the relative expression level of SPART in gastric cancer tissues was 2.78±0.82， which was lower than that in normal gastric tissues （3.84±1.22）， with a statistically significant difference （t=-3.80， P<0.001）. Immunohistochemical staining data from the Human Protein Atlas database showed that among the 23 gastric cancer tissue samples， SPART was lowly expressed in 16 cases （69.57%）， moderately expressed in 7 cases （30.43%）， and had no high expression （0）. Western blotting analysis showed that the relative expression level of SPART protein in gastric cancer tissues in clinical samples was 0.56 （0.68）， which was lower than that in adjacent tissues 1.38 （1.05）， with a statistically significant difference （U=4.00， P=0.016）. CCK-8 assay results showed that at 48 h， the absorbance （A） values of the control and experimental groups of AGS cells were 0.52±0.01 and 0.41±0.01， respectively； those of the control and experimental groups of NUGC3 cells were 0.67±0.02 and 0.51±0.01， respectively， with statistically significant differences （t=13.47， P<0.001； t=12.39， P<0.001）. At 72 h， the A values of the control and experimental groups of AGS cells were 0.82±0.01 and 0.67±0.01， respectively； those of the control and experimental groups of NUGC3 cells were 1.05±0.01 and 0.78±0.02， respectively， with statistically significant differences （t=18.37， P<0.001； t=20.91， P<0.001）. At 96 h， the A values of the control and experimental groups of AGS cells were 1.04±0.03 and 0.74±0.01， respectively； those of the control and experimental groups of NUGC3 cells were 1.80±0.02 and 1.41±0.08， respectively， with statistically significant differences （t=16.43， P<0.001； t=8.19， P=0.004）. Transwell assay revealed that the numbers of migrating cells in the AGS and NUGC3 cell experimental groups were 500±17 and 276±21， respectively， which were less than those in the control groups （771±33 and 327±12）， with statistically significant differences （t=-12.65， P<0.001； t=-3.65， P=0.030）. Western blotting assay showed that， in the control and experimental groups of AGS cells， the expression levels of LC3 protein were 0.47±0.08 and 1.06±0.32， respectively， and the expression levels of LAMP2 protein were 0.65±0.03 and 0.98±0.11， respectively， the expression levels of PLIN3 protein were 0.82±0.15 and 0.43±0.10， respectively， all with statistically significant differences （t=3.11， P=0.040； t=5.02， P=0.007； t=3.75， P=0.020）. In the control and experimental groups of NUGC3 cells， the expression levels of LC3 protein were 0.89±0.04 and 1.45±0.16， respectively， and the expression levels of LAMP2 protein were 0.68±0.18 and 1.22±0.22， respectively， the expression levels of PLIN3 protein were 1.71±0.45 and 0.35±0.06， respectively， all with statistically significant differences （t=5.89， P=0.004； t=3.29， P=0.030； t=5.19， P=0.007）. Immunofluorescence assay showed that， the lipid droplet levels in the AGS cell control group and the experimental group were 1.08±0.09 and 0.32±0.03， respectively， and the LC3 levels were 0.40±0.01 and 0.88±0.15， respectively， with statistically significant differences （t=13.88， P<0.001； t=-5.53， P=0.033）. Conclusions SPART is lowly expressed in gastric cancer， and its overexpression can inhibit gastric cancer cell proliferation and migration while activating lipophagy.

Key words: Stomach neoplasms, Cell proliferation, Cell movement, Lipophagy, SPART

来比江·吾斯曼, 宋钉町, 张文斌. SPART通过脂噬对胃癌细胞增殖和迁移能力的影响[J]. 国际肿瘤学杂志, 2026, 53(2): 73-78.

Laibijiang Wusiman, Song Dingding, Zhang Wenbin. Effect of SPART on the proliferation and migration capabilities of gastric cancer cell through lipophagy[J]. Journal of International Oncology, 2026, 53(2): 73-78.

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