TSR2 overexpression inhibits proliferation and invasion of gastric cancer cells by downregulating the PI3K/AKT signaling pathway

doi:10.12122/j.issn.1673-4254.2024.05.13

Abstract

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

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.

Figures/Tables 8

References 38

1	Mülder DT, Hahn AI, Huang RJ, et al. Prevalence of gastric precursor lesions in countries with differential gastric cancer burden: a systematic review and meta-analysis[J]. Clin Gastroenterol Hepatol, 2024: S1542-S3565(24)00227-1. DOI: 10.1016/j.cgh.2024.02.023
2	Bray F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68(6): 394-424. DOI: 10.3322/caac.21492
3	Digklia A, Wagner AD. Advanced gastric cancer: current treatment landscape and future perspectives[J]. World J Gastroenterol, 2016, 22(8): 2403-14. DOI: 10.3748/wjg.v22.i8.2403
4	Smyth EC, Nilsson M, Grabsch HI, et al. Gastric cancer[J]. Lancet, 2020, 396(10251): 635-48. DOI: 10.1016/s0140-6736(20)31288-5
5	朱正纲. 胃癌外科综合治疗的若干进展与展望[J]. 外科理论与实践, 2023, 28(1): 1-6.
6	Song ZY, Wu YY, Yang JB, et al. Progress in the treatment of advanced gastric cancer[J]. Tumour Biol, 2017, 39(7): 101042831771462. DOI: 10.1177/1010428317714626
7	Brisinda G, Chiarello MM, Crocco A, et al. Postoperative mortality and morbidity after D2 lymphadenectomy for gastric cancer: a retrospective cohort study[J]. World J Gastroenterol, 2022, 28(3): 381-98. DOI: 10.3748/wjg.v28.i3.381
8	Feng F, Tian YZ, Xu GH, et al. Diagnostic and prognostic value of CEA, CA19-9, AFP and CA125 for early gastric cancer[J]. BMC Cancer, 2017, 17(1): 737. DOI: 10.1186/s12885-017-3738-y
9	de Manzoni G, Yang HK. Gastric cancer surgery[J]. Updat Surg, 2018, 70(2):155. DOI: 10.1007/s13304-018-0551-3
10	贾祺, 咸小红, 李泱润, 等. MAGE-A家族在胃癌中作用的研究进展[J]. 中南大学学报: 医学版, 2023, 48(2): 260-7.
11	Zhao QF, Cao L, Guan LL, et al. Immunotherapy for gastric cancer: dilemmas and prospect[J]. Brief Funct Genomics, 2019, 18(2): 107-12. DOI: 10.1093/bfgp/ely019
12	Zheng CH, Xu YC, Zhao G, et al. Outcomes of laparoscopic total gastrectomy combined with spleen-preserving hilar lymphadenectomy for locally advanced proximal gastric cancer: a nonrandomized clinical trial[J]. JAMA Netw Open, 2021, 4(12): e2139992. DOI: 10.1001/jamanetworkopen.2021.39992
13	Lin XL, Han T, Xia Q, et al. CHPF promotes gastric cancer tumorigenesis through the activation of E2F1[J]. Cell Death Dis, 2021, 12(10): 876. DOI: 10.1038/s41419-021-04148-y
14	Feng YY, Zhao M, Wang LJ, et al. The heterogeneity of signaling pathways and drug responses in intrahepatic cholangiocarcinoma with distinct genetic mutations[J]. Cell Death Dis, 2024, 15(1): 34. DOI: 10.1038/s41419-023-06406-7
15	He HJ, Bing H, Liu GJ. TSR2 Induces laryngeal cancer cell apoptosis through inhibiting NF‑κB signaling pathway[J]. Laryngoscope, 2018, 128(4): E130-4. DOI: 10.1002/lary.27035
16	He HJ, Zhu D, Sun J, et al. The Novel protein TSR2 inhibits the transcriptional activity of nuclear factor‑κB and induces apoptosis[J]. Mol Biol, 2011, 45(3): 451-7. DOI: 10.1134/s0026893311020099
17	Zhao QX, Rangan R, Weng SN, et al. Inhibition of ribosome biogenesis in the epidermis is sufficient to trigger organism-wide growth quiescence independently of nutritional status in C. elegans[J]. PLoS Biol, 2023, 21(8): e3002276. DOI: 10.1371/journal.pbio.3002276
18	Bu DC, Luo HT, Huo PP, et al. KOBAS-i: intelligent prioritization and exploratory visualization of biological functions for gene enrichment analysis[J]. Nucleic Acids Res, 2021, 49(W1): W317-25. DOI: 10.1093/nar/gkab447
19	Sun XZ, Chen PX, Chen X, et al. KIF4A enhanced cell proliferation and migration via Hippo signaling and predicted a poor prognosis in esophageal squamous cell carcinoma[J]. Thorac Cancer, 2021, 12(4): 512-24. DOI: 10.1111/1759-7714.13787
20	Thrift AP, El-Serag HB. Burden of gastric cancer[J]. Clin Gastroenterol Hepatol, 2020, 18(3): 534-42. DOI: 10.1016/j.cgh.2019.07.045
21	Guggenheim DE, Shah MA. Gastric cancer epidemiology and risk factors[J]. J Surg Oncol, 2013, 107(3): 230-6. DOI: 10.1002/jso.23262
22	Karimi P, Islami F, Anandasabapathy S, et al. Gastric cancer: descriptive epidemiology, risk factors, screening, and prevention[J]. Cancer Epidemiol Biomarkers Prev, 2014, 23(5): 700-13. DOI: 10.1158/1055-9965.epi-13-1057
23	Maddineni G, Xie JJ, Brahmbhatt B, et al. Diet and carcinogenesis of gastric cancer[J]. Curr Opin Gastroenterol, 2022, 38(6): 588-91. DOI: 10.1097/mog.0000000000000875
24	Yusefi AR, Bagheri Lankarani K, Bastani P, et al. Risk factors for gastric cancer: a systematic review[J]. Asian Pac J Cancer Prev, 2018, 19(3): 591-603.
25	Ansari S, Gantuya B, Tuan VP, et al. Diffuse gastric cancer: a summary of analogous contributing factors for its molecular pathogenicity[J]. Int J Mol Sci, 2018, 19(8): 2424. DOI: 10.3390/ijms19082424
26	Guo ZJ, Guo L. Abnormal activation of RFC3, A YAP1/TEAD downstream target, promotes gastric cancer progression[J]. Int J Clin Oncol, 2024, 29(4): 442-55. DOI: 10.1007/s10147-024-02478-3
27	Rong L, Li ZD, Leng X, et al. Salidroside induces apoptosis and protective autophagy in human gastric cancer AGS cells through the PI3K/Akt/mTOR pathway[J]. Biomed Pharmacother, 2020, 122: 109726. DOI: 10.1016/j.biopha.2019.109726
28	Zanotelli MR, Zhang J, Reinhart-King CA. Mechanoresponsive metabolism in cancer cell migration and metastasis[J]. Cell Metab, 2021, 33(7): 1307-21. DOI: 10.1016/j.cmet.2021.04.002
29	Wan GQ, Liu YH, Zhu J, et al. SLFN5 suppresses cancer cell migration and invasion by inhibiting MT1-MMP expression via AKT/GSK-3β/β-catenin pathway[J]. Cell Signal, 2019, 59: 1-12. DOI: 10.1016/j.cellsig.2019.03.004
30	Yang YM, Karbstein K. The chaperone Tsr2 regulates Rps26 release and reincorporation from mature ribosomes to enable a reversible, ribosome-mediated response to stress[J]. Sci Adv, 2022, 8(8): eabl4386. DOI: 10.1126/sciadv.abl4386
31	Yang YM, Jung Y, Abegg D, et al. Chaperone-directed ribosome repair after oxidative damage[J]. Mol Cell, 2023, 83(9): 1527-37, e5. DOI: 10.1016/j.molcel.2023.03.030
32	Fernández-Parejo N, Lorenzo-Martín LF, García-Pedrero JM, et al. VAV2 orchestrates the interplay between regenerative proliferation and ribogenesis in both keratinocytes and oral squamous cell carcinoma[J]. Sci Rep, 2024, 14(1): 4060. DOI: 10.1038/s41598-024-54808-0
33	Zang Y, Ran X, Yuan J, et al. Genomic hallmarks and therapeutic targets of ribosome biogenesis in cancer[J]. Brief Bioinform, 2024, 25(2): bbae023. DOI: 10.1093/bib/bbae023
34	An Y, Xia YC, Wang ZY, et al. Clinical significance of ribosome production factor 2 homolog in hepatocellular carcinoma[J]. Clin Res Hepatol Gastroenterol, 2024, 48(3): 102289. DOI: 10.1016/j.clinre.2024.102289
35	Miricescu D, Totan A, Stanescu-Spinu II, et al. PI3K/AKT/mTOR signaling pathway in breast cancer: from molecular landscape to clinical aspects[J]. Int J Mol Sci, 2020, 22(1): 173. DOI: 10.3390/ijms22010173
36	Sun TS, Bi FF, Liu ZN, et al. TMEM119 facilitates ovarian cancer cell proliferation, invasion, and migration via the PDGFRB/PI3K/AKT signaling pathway[J]. J Transl Med, 2021, 19(1): 111. DOI: 10.1186/s12967-021-02781-x
37	Fresno Vara JA, Casado E, de Castro J, et al. PI3K/Akt signalling pathway and cancer[J]. Cancer Treat Rev, 2004, 30(2): 193-204. DOI: 10.1016/j.ctrv.2003.07.007
38	Maiello D, Varone M, Vicidomini R, et al. Dyskerin downregulation can induce ER stress and promote autophagy via AKT-mTOR signaling deregulation[J]. Biomedicines, 2022, 10(5): 1092. DOI: 10.3390/biomedicines10051092

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%)

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%)

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

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

[1]	Yan WANG, Yuqing RUAN, Can CUI, Xiu WANG. Jiaotaiwan improves brain glucose metabolism in a mouse model of Alzheimer's disease by activating the PI3K/AKT signaling pathway [J]. Journal of Southern Medical University, 2024, 44(5): 894-903.
[2]	Yuanyuan WANG, Teng CHEN, Xiaofan CONG, Yiran LI, Rui CHEN, Pei ZHANG, Xiaojin SUN, Surong ZHAO. Pristimerin enhances cisplatin-induced apoptosis in nasopharyngeal carcinoma cells via ROS-mediated deactivation of the PI3K/AKT signaling pathway [J]. Journal of Southern Medical University, 2024, 44(5): 904-912.
[3]	HUANG Qiuhu, ZHOU Jian, WANG Zizhen, YANG Kun, CHEN Zhenggang. MiR-26-3p regulates proliferation, migration, invasion and apoptosis of glioma cells by targeting CREB1 [J]. Journal of Southern Medical University, 2024, 44(3): 578-584.
[4]	LIANG Yihao, LAI Yingjun, YUAN Yanwen, YUAN Wei, ZHANG Xibo, ZHANG Bashan, LU Zhifeng. Screening of differentially expressed genes in gastric cancer based on GEO database and function and pathway enrichment analysis [J]. Journal of Southern Medical University, 2024, 44(3): 605-616.
[5]	ZHU Jin, OUYANG Xin, LIU Yu, QIAN Yemei, XIA Bin, SHI Yanan, YU Lifu. MiR-132-3p negatively regulates CAMTA1 to promote Schwann cell proliferation and migration and alleviates I-125 seeds-induced exacerbation of facial nerve injury in rats [J]. Journal of Southern Medical University, 2024, 44(3): 571-577.
[6]	SHEN Mengdi, ZHAO Na, DENG Xiaojing, DENG Min. High expression of COX6B2 in gastric cancer is associated with poor long-term prognosis and promotes cell proliferation and cell cycle progression by inhibiting p53 signaling [J]. Journal of Southern Medical University, 2024, 44(2): 289-297.
[7]	WU Guangyang, SONG Tianli, TANG Lang, WANG Yiming, LIU Xu, HUANG Sheng. Total saponins of Panax japonicus alleviates CCl4-induced acute liver injury in rats by regulating the PI3K/AktNF-κB signaling pathway [J]. Journal of Southern Medical University, 2024, 44(2): 244-251.
[8]	ZHANG Nuo, ZHANG Zhen, ZHANG Yulu, SONG Xue, ZHANG Xiaofeng, LI Jing, ZUO Lugen, HU Jianguo. PCID2 is highly expressed in gastric cancer and affects the prognosis by regulating cancer cell cycle and proliferation [J]. Journal of Southern Medical University, 2024, 44(2): 324-332.
[9]	ZHANG Wenjing, ZHANG Nuo, YANG Zi, ZHANG Xiaofeng, SUN Aofei, WANG Lian, SONG Xue, GENG Zhijun, LI Jing, HU Jianguo. Overexpression of BZW1 promotes invasion and metastasis of gastric cancer cells by regulating Wnt/β-catenin signaling and promoting epithelial-mesenchymal transition [J]. Journal of Southern Medical University, 2024, 44(2): 354-362.
[10]	ZHONG Weixiong, LIANG Fangrong, YANG Ruimeng, ZHEN Xin. Prediction of microvascular invasion in hepatocellular carcinoma based on multi-phase dynamic enhanced CT radiomics feature and multi-classifier hierarchical fusion model [J]. Journal of Southern Medical University, 2024, 44(2): 260-269.
[11]	DUAN Ting, ZHANG Zhen, SHI Jinran, XIAO Linyu, YANG Jingjing, YIN Lixia, ZHANG Xiaofeng, GENG Zhijun, LU Guoyu. High expression of CPNE3 correlates with poor long-term prognosis of gastric cancer by inhibiting cell apoptosis via activating PI3K/AKT signaling [J]. Journal of Southern Medical University, 2024, 44(1): 129-137.
[12]	FENG Wen, LAI Yuexing, WANG Jing, XU Ping. Long non-coding RNA ABHD11-AS1 promotes glycolysis in gastric cancer cells to accelerate tumor progression [J]. Journal of Southern Medical University, 2023, 43(9): 1485-1492.
[13]	LIU Xuerou, YANG Yumei, CAI Hui, ZHANG Yaoshuai, FAN Fangtian, LI Xian, LI Shanshan. Aumolertinib inhibits proliferation, invasion and migration and promotes apoptosis of neuroblastoma cells by downregulating MMP2 and MMP9 expression [J]. Journal of Southern Medical University, 2023, 43(9): 1493-1499.
[14]	WANG Lian, XIA Yongsheng, ZHANG Zhen, LIU Xinyue, SHI Jinran, WANG Yueyue, LI Jing, ZHNAG Xiaofeng, GENG Zhijun, SONG Xue, ZUO Lugen. High expression of MRPL13 promotes cell cycle progression and proliferation of gastric cancer cells by inhibiting p53 signaling to affect long-term prognosis [J]. Journal of Southern Medical University, 2023, 43(9): 1558-1566.
[15]	YU Zhengtao, LI Jiameng, JIANG Junwen, LI You, LIN Long, XIA Ying, WANG Lei. miRNA-128-3p inhibits malignant behavior of glioma cells by downregulating KLHDC8A expression [J]. Journal of Southern Medical University, 2023, 43(9): 1447-1459.