过表达TSR2通过下调PI3K/AKT信号通路抑制胃癌细胞的增殖和侵袭

doi:10.12122/j.issn.1673-4254.2024.05.13

摘要/Abstract

摘要：

目的探讨TSR2核糖体成熟因子在胃癌中的表达情况及其与胃癌恶性演进的相关性，并分析其潜在的作用机制。方法纳入105例胃癌患者资料，分析TSR2在胃癌组织中表达水平及其对胃癌恶性进展、术后5年生存率的影响；GO及KEGG富集分析预测TSR2的生物学功能及可能的作用机制；通过慢病毒转染技术上调和下调TSR2在胃癌细胞系（MGC-803）的表达水平，并采用CCK-8、Transwell评估其对MGC-803细胞增殖、侵袭及迁移的影响；Western blot检测p-PI3K、p-AKT表达。结果 TSR2在胃癌组织的表达水平显著低于癌旁组织（P<0.001），且TSR2的表达水平与CEA、CA19-9、T分期及N分期相关（P<0.05）。单因素联合多因素分析显示，TSR2低表达（P=0.020）、CEA≥5 μg/L（P=0.021）、CA19-9≥37 kU/L（P=0.001）、T₃~T₄分期（P=0.039）和N₂~N₃分期（P=0.027）是独立影响胃癌患者施行根治术后5年生存率的风险因子。生存分析结果显示，TSR2表达水平与胃癌患者术后5年生存率呈正相关（P<0.001）。生物信息学富集分析预测TSR2的功能可能与PI3K/AKT信号通路相关。CCK-8和Transwell实验结果显示，上调TSR2可抑制胃癌细胞的增殖、迁移和侵袭（P<0.05），下调则反之（P<0.05）。Western blot结果显示过表达TSR2可下调胃癌细胞中磷脂肌醇3激酶（PI3K）和蛋白激酶B（AKT）的磷酸化，敲低则反之（P<0.05）。结论 TSR2在胃癌组织中低表达并影响患者预后，其可能与下调PI3K/AKT信号通路抑制胃癌细胞的增殖、侵袭与迁移有关。

关键词: 胃癌, TSR2核糖体成熟因子, PI3K/AKT, 增殖, 侵袭

Abstract:

Objective To investigate the expression of TSR2 in gastric cancer and explore its correlation with progression of gastric cancer and the possible mechanism. Methods We retrospectively analyzed TSR2 expression in clinical specimens from 105 gastric cancer patients and the impact of TSR2 expression level on disease progression and 5-year postoperative survival of the patients. GO and KEGG enrichment analyses were used to predict the biological functions and mechanisms of TSR2. In gastric cancer MGC-803 cells with lentivirus-mediated TSR2 overexpression or knockdown, the changes in cell proliferation, invasion, and migration were assessed with CCK-8 and Transwell assays, and the expressions of p-PI3K and p-AKT were detected using Western blotting. Results TSR2 expression was significantly lower in gastric cancer tissues than in the adjacent tissues with significant correlations with CEA level, CA19-9 level, and T and N staging (P<0.05). A low TSR2 expression, CEA≥5 μg/L, CA19-9≥37 kU/L, T₃-T₄ stages, and N₂-N₃ staged were identified as independent risk factors affecting 5-year survival rate of the patients following radical surgery (P<0.05), and a high TSR2 expression was associated with a higher 5-year survival rate of the patients (P<0.001). Bioinformatics analysis suggested the functional involvement of TSR2 with the PI3K/AKT signaling pathway. MGC-803 cells overexpressing TSR2 showed significantly lowered proliferation, migration, and invasion capacities (P<0.05), while TSR2 knockdown produced the opposite effects (P<0.05). Western blotting showed that TSR2 overexpression reduced the phosphorylation of PI3K and AKT, and TSR2 knockdown caused the opposite changes in MGC-803 cells (P<0.05). Conclusion TSR2 is lowly expressed in gastric cancer tissues to adversely affect the patients' prognosis, and its overexpression inhibits gastric cancer cell proliferation, invasion, and migration possibly by downregulating the PI3K/AKT pathway.

Key words: gastric cancer, TSR2 ribosome maturation factor, PI3K/AKT, proliferation, invasion

夏勇生, 王炼, 陈孝华, 张雨路, 孙奥飞, 陈德利. 过表达TSR2通过下调PI3K/AKT信号通路抑制胃癌细胞的增殖和侵袭[J]. 南方医科大学学报, 2024, 44(5): 913-919.

Yongsheng XIA, Lian WANG, Xiaohua CHEN, Yulu ZHANG, Aofei SUN, Deli CHEN. TSR2 overexpression inhibits proliferation and invasion of gastric cancer cells by downregulating the PI3K/AKT signaling pathway[J]. Journal of Southern Medical University, 2024, 44(5): 913-919.

图/表 8

图1 TSR2在胃癌及癌旁组织中的表达情况

Fig.1 Expression of TSR2 in gastric cancer and adjacent tissues. A: TSR2 is lowly expressed in gastric cancer tissue. B: TSR2 is highly expressed in adjacent tissue. C: Relative IOD value of TSR2 (n=105). *P<0.05 vs adjacent tissues.

表1 胃癌组织中TSR2的表达量与恶性进展参数的关系

Tab.1 Correlation between TSR2 expression and progression of gastric cancer [n (%)]

Factors	n	TSR2 expression		χ ²	P
Factors	n	Low (n=53)	High (n=52)	χ ²	P
Gender				0.023	0.880
Male	74	37 (50.0%)	37 (50.0%)
Female	31	16 (51.6%)	15 (48.4%)
Age (year)				0.015	0.903
<60	41	21 (51.2%)	20 (48.8%)
≥60	64	32 (50.0%)	32 (50.0%)
CEA (μg/L)				6.940	0.008
<5	49	18 (36.7%)	31 (63.3%)
≥5	56	35 (62.5%)	21 (37.5%)
CA19-9 (kU/L)				17.720	<0.001
<37	47	13 (27.7%)	34 (72.3%)
≥37	58	40 (69.0%)	18 (31.0%)
Tumor size (cm)				2.134	0.144
<5	49	21 (42.9%)	28 (57.1%)
≥5	56	32 (57.1%)	24 (42.9%)
Histological type				1.177	0.278
Adenocarcinoma	66	36 (54.5%)	30 (45.5%)
Other	39	17 (43.6%)	22 (56.4%)
T stage				11.675	0.001
1-2	53	18 (34.0%)	35 (66.0%)
3-4	52	35 (67.3%)	17 (32.7%)
N stage				13.414	<0.001
0-1	60	21 (35.0%)	39 (65.0%)
2-3	45	32 (71.1%)	13 (28.9%)

表2 影响胃癌患者预后的危险因素

Tab.2 Risk factors affecting prognosis of patients with gastric cancer

Characteristic	Univariate analysis		Multivariate analysis
Characteristic	Log rank χ²	P	HR	95% CI	P
Gender (male vs female)	1.451	0.228	-	-	-
Age (≥60 years vs <60 years)	1.356	0.244	-	-	-
TSR2 expression (low vs high)	31.840	<0.001	0.418	0.201-0.871	0.020
CEA (≥5μg/L vs <5 μg/L)	18.367	<0.001	2.173	1.126-4.192	0.021
CA19-9 (≥37 kU/L vs <37 kU/L)	25.491	<0.001	3.144	1.559-6.342	0.001
Tumor size (≥5 cm vs <5 cm)	2.163	0.141	-	-	-
Histological type(adenocarcinoma vs other)	0.001	0.982	-	-	-
T stage (T₃-T₄vs T₁-T₂)	21.658	<0.001	1.991	1.034-3.832	0.039
N stage (N₂-N₃vs N₀-N₁)	26.362	<0.001	2.106	1.090-4.066	0.027

图2 胃癌组织中TSR2表达水平对患者术后5年生存率的影响

Fig.2 Influence of TSR2 expression level in gastric cancer tissue on 5-year survival rate of the patients after surgery.

图3 TSR2的KEGG和GO富集分析

Fig.3 KEGG and GO enrichment analysis of TSR2. A: KEGG analysis of TSR2 in gastric cancer. B: GO analysis of TSR2 in gastric cancer.

图4 TSR2对MGC-803细胞增殖能力的影响

Fig.4 Effect of TSR2 overexpression and knockdown on proliferation of MGC-803 cells. A, B: Lentivirus-mediated TSR2 overexpression and knockdown in MGC803 cells. C: TSR2 overexpression inhibits proliferation of gastric cancer cells (n=3). LV: Overexpression; Si: SiRNA. *P<0.05 vs Control.

图5 TSR2对MGC-803细胞迁移和侵袭能力的影响

Fig.5 Effects of TSR2 overexpression and knockdown on migration (A) and invasion (B) of MGC-803 cells detected by Transwell assay (n=3). *P<0.05 vs Control.

图6 TSR2调控MGC-803细胞中的PI3K/AKT信号通路

Fig.6 Western blotting for assessing the effect of TSR2 overexpression and knockdown on expression of p-PI3K and p-AKT in MGC803 cells (n=3). *P<0.05 vs Control.

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