Expression of SKA3 protein in cholangiocarcinoma tissues and its effect on the function of cholangiocarcinoma SSP-25 cells in vitro and PI3K-AKT signaling pathway
10.3760/cma.j.cn115355-20240221-00082
- VernacularTitle:SKA3蛋白在胆管癌组织中的表达及其体外对胆管癌SSP-25细胞功能和PI3K-AKT信号通路的影响
- Author:
Dong WANG
1
;
Yixi ZHANG
;
Chengjian GUAN
;
Xiaodong ZHANG
;
Huajun LIN
;
Wei GUO
Author Information
1. 首都医科大学附属北京友谊医院普通外科，北京 100050
- Keywords:
Bile duct neoplasms;
SKA3 protein;
Cell proliferation;
Neoplasm invasiveness;
PI3K-AKT signaling pathway
- From:
Cancer Research and Clinic
2024;36(5):335-340
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the expression and clinical significance of SKA3 protein in cholangiocarcinoma, and the effect of interfering SKA3 expression in vitro on the proliferation, invasion, and migration of cholangiocarcinoma SSP-25 cells, as well as its possible mechanism.Methods:The clinicopathological data, cancer tissues, and paracancerous tissues from 172 patients with distal cholangiocarcinoma in Beijing Friendship Hospital, Capital Medical University between January 2015 and December 2020 were retrospectively collected. Immunohistochemical method was used to detect the expression level of SKA3 protein in cancer tissues and paracancerous tissues. Transfection of SKA3 small interfering RNA (siRNA) into cholangiocarcinoma SSP-25 cells was used as si-SKA3 group, and the untreated SSP-25 cells were used as the control group. Cell immunofluorescence staining and Western blot were used to detect the transfection effect; CCK-8 method and cell colony formation experiment were used to observe changes in cell proliferation; cell scratch assay was used to monitor cell invasion; Western blot was used to detect the expression of PI3K-AKT signaling pathway related proteins.Results:Among 172 patients with cholangiocarcinoma, there were 116 males and 56 females; the age of 54 cases was under 60 years, and age of 118 cases was equal to or more than 60 years. The positive rate of SKA3 protein in cholangiocarcinoma tissues was higher than that in paracancerous tissues [78.49% (135/172) vs. 13.95% (24/172)], and the difference was statistically significant ( χ2 = 42.78, P < 0.01). The positive rate of SKA3 protein in cancer tissues of cholangiocarcinoma patients with nerve invasion [84.35% (124/147) vs. 44.00% (11/25)] and lymph node metastasis [88.78% (87/98) vs. 64.86% (48/74)] was higher than that of patients without nerve invasion and without lymph node metastasis, and the differences were statistically significant (all P < 0.05). There were no statistically significant differences in the positive rate of SKA3 protein in cancer tissues of patients stratified by age, gender, tumor diameter, TNM stage, and tumor differentiation (all P > 0.05). The CCK-8 method showed that after 72 h of cultivation, the proliferation ability of SSP-25 cells in the si-SKA3 group (expressed as absorbance value at 450 nm) was lower than that in the control group (0.56±0.05 vs. 0.83±0.06), and the difference was statistically significant ( t = 3.06, P = 0.06). After 2 weeks of cultivation, the colony formation experiment showed that the number of colony formation of SSP-25 cells in the si-SKA3 group was lower than that in the control group. After 24 h of cultivation, the scratch healing rates of SSP-25 cells in the si-SKA3 group and the control group were (31±6) % and (72±5)%, respectively, and the difference was statistically significant ( t = 5.63, P = 0.013).Western blot analysis showed that the relative expression levels of p-PI3K and p-AKT proteins in the PI3K-AKT signaling pathway were lower than those in the control group, and the difference was statistically significant (all P < 0.05). Conclusions:SKA3 protein is highly expressed in cholangiocarcinoma tissues, and may related to nerve invasion and lymph node metastasis. Interfering SKA3 expression can inhibit the proliferation and invasion of cholangiocarcinoma SSP-25 cells, and its mechanism may be related to the inhibition of the PI3K-AKT signaling pathway.
