沉默TRPV2基因和大麻二酚对口腔鳞状细胞癌CAL-27细胞生物学行为的影响

doi:10.13481/j.1671-587X.20260115

摘要/Abstract

摘要：

目的探讨瞬时受体电位香草酸通道2（TRPV2）对口腔鳞状细胞癌（OSCC）细胞增殖、迁移和侵袭的影响以及大麻二酚（CBD）通过TRPV2通道对OSCC细胞生物学行为的影响，并阐明其相关抗肿瘤机制。方法利用脂质体法将小干扰RNA（siRNA）片段转染至CAL-27细胞，分为si-NC组（转染无相关性的si-NC）和si-TRPV2组（转染沉默TRPV2基因的特异性siRNA）；不同剂量CBD培养CAL-27细胞，分为0、10、20和40 μmol·L?1 CBD组，采用实时荧光定量PCR（RT-qPCR）法和Western blotting法检测各组细胞中TRPV2 mRNA和蛋白表达水平，采用细胞计数试剂盒8（CCK-8）法、细胞划痕愈合实验和Transwell小室实验分别检测各组CAL-27细胞增殖活性、细胞划痕愈合率和侵袭细胞数。结果 RT-qPCR法和Western blotting法，si-NC组CAL-27细胞中TRPV2 mRNA和蛋白表达水平均高于si-TRPV2组（P<0.01）。CCK-8法，与si-NC组比较，si-TRPV2组转染后48和72 h细胞增殖活性均明显降低（P<0.01）。细胞划痕愈合实验，与si-NC组比较，si-TRPV2组24 h CAL-27细胞划痕愈合率明显降低（P<0.01）。Transwell小室实验，与si-NC组比较，si-TRPV2组侵袭细胞数减少（P<0.01）；RT-qPCR法和Western blotting法，与0 μmol·L?1 CBD组比较，10、20和40 μmol·L?1 CBD组CAL-27细胞中TRPV2 mRNA及蛋白表达水平均降低（P<0.05或P<0.01）；CCK-8法，与0 μmol·L?1 CBD组比较，20和40 μmol·L?1 CBD组药物培养后48、72和96 h细胞存活率均明显降低（P<0.01）；细胞划痕愈合实验，CBD培养12和24 h，与0 μmol·L?1 CBD组比较，10、20和 40 μmol·L?1 CBD组CAL-27细胞划痕愈合率均明显降低（P<0.01）；Transwell小室实验，与0 μmol·L?1 CBD组比较，40 μmol·L^-1 CBD组侵袭细胞数减少（P<0.01）。结论沉默TRPV2基因和CBD处理均能抑制CAL-27细胞增殖、迁移和侵袭能力，CBD能降低CAL-27细胞中TRPV2基因和蛋白表达。

关键词: 瞬时受体电位香草酸通道2, 口腔鳞状细胞癌, 大麻二酚, 细胞迁移, 细胞侵袭

Abstract:

Objective To discuss the effects of transient receptor potential vanilloid channel 2 （TRPV2） on the proliferation， migration， and invasion of oral squamous cell carcinoma （OSCC） cells and the effect of cannabidiol （CBD） on the biological behaviors of OSCC cells through the TRPV2 channel， and to clarify its related anti-tumor mechanism. Methods The siRNA fragments were transfected into the CAL-27 cells using the liposome method， and the cells were divided into si-NC group （transfected with non-related si-NC） and si-TRPV2 group （transfected with specific siRNA silencing TRPV2 gene）. The CAL-27 cells were cultured with different doses of CBD and divided into 0， 10， 20， and 40 μmol·L^-1 CBD groups. Real-time fluorescence quantitative PCR （RT-qPCR） method and Western blotting method were used to detect the expression levels of TRPV2 mRNA and protein in the cells in various groups； cell counting kit-8 （CCK-8） method， cell scratch healing assay， and Transwell chamber assay were used to detect the proliferation activity， scratch healing rate， and number of invasion CAL-27 cells in various groups， respectively. Results The RT-qPCR method and Western blotting method results showed that the expression levels of TRPV2 mRNA and protein in the CAL-27 cells in si-NC group were higher than those in si-TRPV2 group （P<0.01）. The CCK-8 assay results showed that compared with si-NC group， the proliferation activities of the cells in si-TRPV2 group at 48 and 72 h after transfection were significantly decreased （P<0.01）. The cell scratch healing assay results showed that compared with si-NC group， the scratch healing rate of the CAL-27 cells in si-TRPV2 group at 24 h was significantly decreased （P<0.01）. The Transwell chamber assay results showed that compared with si-NC group， the number of invasion cells in si-TRPV2 group was significantly decreased （P<0.01）. The RT-qPCR method and Western blotting method results showed that compared with 0 μmol·L^-1 CBD group， the expression levels of TRPV2 mRNA and protein in the CAL-27 cells in 10， 20， and 40 μmol·L^-1 CBD groups were decreased （P<0.05 or P<0.01）. The CCK-8 assay results showed that compared with 0 μmol·L^-1 CBD group， the cell survival rates of the cells in 20 and 40 μmol·L^-1 CBD groups at 48， 72， and 96 h after drug culture were significantly decreased （P<0.01）. The cell scratch healing assay results showed that after CBD culture for 12 and 24 h， compared with 0 μmol·L^-1 CBD group， the scratch healing rates of the CAL-27 cells in 10， 20， and 40 μmol·L^-1 CBD groups were significantly decreased （P<0.01）. The Transwell chamber assay results showed that compared with 0 μmol·L^-1 CBD group， the number of invasion cells in 40 μmol·L^-1 CBD group was significantly decreased （P<0.01）. Conclusion Silencing the TRPV2 gene and CBD treatment can both inhibit the proliferation， migration， and invasion abilities of CAL-27 cells， and CBD can reduce the expression of TRPV2 gene and protein in CAL-27 cells.

Key words: Transient receptor potential vanilloid channel 2, Oral squamous cell carcinoma, Cannabidiol, Cell migration, Cell invasion

中图分类号:

R739.8

丁云杉,代海涛,陈敏,郝小慧,周肖,仵楠. 沉默TRPV2基因和大麻二酚对口腔鳞状细胞癌CAL-27细胞生物学行为的影响[J]. 吉林大学学报(医学版), 2026, 52(1): 143-151.

Yunshan DING,Haitao DAI,Min CHEN,Xiaohui HAO,Xiao ZHOU,Nan WU. Effect of silencing TRPV2 gene and cannabidiol on biological behaviors of oral squamous cell carcinoma CAL-27 cells[J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 143-151.

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Group	Survival rate
Group	（t/h） 24	48	72	96
CBD （μmol·L^-1）
0	100.00±11.84	100.00±6.61	100.00±7.54	100.00±12.08
10	96.98±7.41	108.70±17.11	118.50±4.76^*	100.80±15.30
20	35.35±1.91^*	17.06±15.87^*	4.36±0.19^*	3.98±0.64^*
40	16.95±0.94^*	10.29±5.07^*	3.99±1.26^*	2.84±1.65^*