Gli gene and protein expression in hepatocellular carcinoma and the relationship with prognosis

doi:10.11958/20240977

Abstract

Abstract:

Objective To investigate the expression patterns of Gli1, Gli2 and Gli3 genes and proteins in hepatocellular carcinoma (HCC) tissue and their correlation with prognosis. Methods Gli gene expression in 315 HCC samples were analyzed using the cBioportal database, and Gli protein expression was examined in 40 HCC and other liver tissue samples through immunohistochemistry. Results Gli gene alterations were found in 57 cases (18.1%) of HCC samples, with alteration frequencies of 19 cases (6.0%), 13 cases (4.1%) and 25 cases (7.9%) for Gli1, Gli2 and Gli3, respectively. Gli1 showed significant co-occurrence with both Gli2 and Gli3 (P<0.05), while Gli2 and Gli3 exhibited a tendency toward mutual exclusivity. Increased Gli1 gene expression significantly correlated with decreased disease-free survival (P<0.01). Immunohistochemical analysis demonstrated that Gli1 protein expression was significantly higher in HCC tissue than that in normal liver tissue, adjacent tissue, hepatitis and cirrhotic tissue (P<0.05). In HCC tissue, strong positive correlation was observed between the three Gli proteins (Gli1 vs. Gli2: r_s=0.796; Gli1 vs. Gli3: r_s=0.759; Gli2 vs. Gli3:r_s=0.891, all P<0.01). High expression levels of all three Gli proteins were detected in 53.3% (8/15) of HCC samples, compared to only 11.8% (2/17) in non-cancerous liver tissue. Conclusion Gli1, Gli2 and Gli3 proteins commonly exhibit synergistic high expression and significant positive correlation in HCC tissue, with Gli1 showing a special expression pattern of independent high expression.

Key words: liver neoplasms, transcription factors, gene expression, Gli, cBioportal gene database

CLC Number:

R735.7

Figures/Tables 6

Fig.1 Alterations in the expression levels of Gli1, Gli2 and Gli3 genes in hepatocellular carcinoma

Tab.1 Comparison of exclusivity and coexistence between three types of Gli gene

基因A	基因B	共同改变	只有A改变	只有B改变	均无改变	$L o g 2 O R$	P	趋势
Gli1	Gli3	7	15	21	305	2.761	<0.001	共存
Gli1	Gli2	3	19	10	316	2.319	0.042	共存
Gli2	Gli3	0	13	28	307	<-3.000	0.329	互斥

Tab.1 Comparison of exclusivity and coexistence between three types of Gli gene

基因A	基因B	共同改变	只有A改变	只有B改变	均无改变	$L o g 2 O R$	P	趋势
Gli1	Gli3	7	15	21	305	2.761	<0.001	共存
Gli1	Gli2	3	19	10	316	2.319	0.042	共存
Gli2	Gli3	0	13	28	307	<-3.000	0.329	互斥

Fig.2 Correlation between Gli gene alterations and overall survival as well as disease-free survival in hepatocellular carcinoma

Fig.3 Expression of Gli1, Gli2 and Gli3 proteins in normal tissue and hepatocellular carcinoma tissue (×200)

Tab.2 Comparison of Gli1, Gli2 and Gli3 expression between normal tissue and diseased liver tissue

组织类型	n	Gli1	Gli2	Gli3
正常肝脏	2	0.00（0.00，0.01）	0.06（0.05，0.07）	0.02（0.01，0.02）
癌旁组织	2	0.01（0.00，0.02）	0.13（0.08，0.16）	0.05（0.04，0.07）
病毒性肝炎	5	0.02（0.00，0.05）	0.12（0.09，0.16）	0.06（0.03，0.07）
肝硬化/ 肝纤维化	8	0.05（0.02，0.09）	0.11（0.06，0.19）	0.05（0.02，0.07）
HCC	15	0.21（0.06，0.50）^abcd	0.09（0.04，0.20）^a	0.06（0.02，0.09）^a
H		25.370^＊＊	9.820^＊	11.730^＊

Fig.4 Co-expression patterns of Gli1, Gli2 and Gli3 proteins in each sample of normal tissue and diseased liver tissue

References 21

[1]	WANG C Y, LI S. Clinical characteristics and prognosis of 2887 patients with hepatocellular carcinoma:a single center 14 years experience from China[J]. Medicine(Baltimore), 2019, 98(4):e14070. doi:10.1097/MD.0000000000014070.
[2]	MACHADO M V, DIEHL A M. Hedgehog signalling in liver pathophysiology[J]. J Hepatol, 2018, 68(3):550-562. doi:10.1016/j.jhep.2017.10.017.
[3]	王雷, 杜媛鲲, 王林, 等. Gli抑制剂GANT61对食管腺癌细胞生长和转移抑制作用的研究[J]. 天津医药, 2016, 44(5):556-559.
	WANG L, DU Y K, WANG L, et al. GANT61 as an inhibitor of Gli inhibits growth and invasion of esophageal adenocarcinoma[J]. Tianjin Med J, 2016, 44(5):556-559. doi:10.11958/20150351.
[4]	HAN W, ALLAM S A, ELSAWA S F. GLI2-mediated inflammation in the tumor microenvironment[J]. Adv Exp Med Biol, 2020, 1263:55-65. doi:10.1007/978-3-030-44518-8_5.
[5]	ZHANG Y X, PHILIP A B. Cellular and molecular mechanisms of Hedgehog signalling[J]. Nat Rev Mol Cell Biol, 2023, 24(9):668-687. doi:10.1038/s41580-023-00591-1.
[6]	YOSHIDA S, KAWAMURA A, AOKI K, et al. Positive regulation of Hedgehog signaling via phosphorylation of GLI2/GLI3 by DYRK2 kinase[J]. Proc Natl Acad Sci U S A, 2024, 121(28):e2320070121. doi:10.1073/pnas.2320070121.
[7]	JENG K S, JENG C J, JENG W J, et al. Sonic Hedgehog signaling pathway as a potential target to inhibit the progression of hepatocellular carcinoma[J]. Oncol Lett, 2019, 18(5):4377-4384. doi:10.3892/ol.2019.10826.
[8]	HARADA K, OHASHI R, NAITO K, et al. Hedgehog signal inhibitor GANT61 inhibits the malignant behavior of undifferentiated hepatocellular carcinoma cells by targeting non-canonical Gli signaling[J]. Int J Mol Sci, 2020, 21(9):3126. doi:10.3390/ijms21093126.
[9]	Cancer Genome Atlas Research Network. Comprehensive and integrative genomic characterization of hepatocellular carcinoma[J]. Cell, 2017, 169(7):1327-1341. doi:10.1016/j.cell.2017.05.046.
[10]	CLARA J A, MONGE C, YANG Y, et al. Targeting signalling pathways and the immune microenvironment of cancer stem cells - a clinical update[J]. Nat Rev Clin Oncol, 2020, 17(4):204-232. doi:10.1038/s41571-019-0293-2.
[11]	MASSAH S, FOO J, LI N, et al. Gli activation by the estrogen receptor in breast cancer cells:Regulation of cancer cell growth by Gli3[J]. Mol Cell Endocrinol, 2021, 522:111136. doi:10.1016/j.mce.2020.111136.
[12]	RODRIGUES M, MIGUITA L, DE ANDRADE N P, et al. GLI3 knockdown decreases stemness,cell proliferation and invasion in oral squamous cell carcinoma[J]. Int J Oncol, 2018, 53(6):2458-2472. doi:10.3892/ijo.2018.4572.
[13]	ZHU H, XIA L, SHEN Q, et al. Differential effects of GLI2 and GLI3 in regulating cervical cancer malignancy in vitro and in vivo[J]. Lab Invest, 2018, 98(11):1384-1396. doi:10.1038/s41374-018-0089-5.
[14]	MATISSEK S J, ELSAWA S F. Gli3:A mediator of genetic diseases, development and cancer[J]. Cell Commun Signal, 2020, 18(1):54. doi:10.22099/mbrc.2024.49044.1915.
[15]	WANG S, WANG Y, XUN X, et al. Hedgehog signaling promotes sorafenib resistance in hepatocellular carcinoma patient-derived organoids[J]. J Exp Clin Cancer Res, 2020, 39(1):22. doi:10.1186/s13046-020-1523-2.
[16]	HU Y, PENG L, ZHUO X, et al. Hedgehog signaling pathway in fibrosis and targeted therapies[J]. Biomolecules, 2024, 14(12):1485. doi:10.3390/biom14121485.
[17]	DING J, YANG Y Y, LI P T, et al. TGF-β1/SMAD3-driven GLI2 isoform expression contributes to aggressive phenotypes of hepatocellular carcinoma[J]. Cancer Lett, 2024, 588:216768. doi:10.1016/j.canlet.2024.216768.
[18]	PIETROBONO S, GAGLIARDI S, STECCA B. Non-canonical Hedgehog signaling pathway in cancer:activation of Gli transcription factors beyond smoothened[J]. Front Genet, 2019, 10:556. doi:10.3389/fgene.2019.00556.
[19]	WU H, WEI M, LI Y, et al. Research progress on the regulation mechanism of key signal pathways affecting the prognosis of glioma[J]. Front Mol Neurosci, 2022, 15:910543. doi:10.3389/fnmol.2022.910543.
[20]	NIEWIADOMSKI P, NIEDZIÓŁKA S M, MARKIEWICZ Ł, et al. Gli proteins:regulation in development and cancer[J]. Cells, 2019, 8(2):147. doi:10.3390/cells8020147.
[21]	UMBERGER P A, OGDEN S K. SPOP and CUL3 modulate the sonic hedgehog signal response through controlled degradation of Gli family transcription factors[J]. Front Cell Dev Biol, 2021, 9:710295. doi:10.3389/fcell.2021.710295.