缺氧对宫颈癌SiHa细胞中Gli1与Sox2表达的影响

doi:10.13481/j.1671-587X.20240217

Abstract

Abstract:

Objective To analyze the expressions of glioma-associated oncogene homolog 1 （Gli1） and sex determining region Y 2 （Sox2） in the cervical cancer cells and their pathological significances， and to discuss the correlation between the expression of Gli1 and the stemness characteristics of the cervical cancer. Methods Immunohistochemical staining was used to detect the expression of Gli1， hypoxia-inducible factor 1-alpha （HIF-1α）， Sox2， sex determining region Y 9 （Sox9）， cluster of differentiation 44 （CD44）， octamer-binding transcription factor 4 （OCT4）， and lysine-specific demethylase 1 （LSD1） in 159 cases of cervical cancer tissues and their correlations were analyzed. Gene Expression Profiling Interactive Analysis （GEPIA） was used to clarify the effect of Gli1 expression on the prognosis of the patients with cervical carcinoma and its correlation with the stem cell markers. The sphere-forming ability and the expression of stem cell markers in the SiHa cells under hypoxic status were detected. Results The immunohistochemical staining results showed that the expression rate of Gli1 in cervical cancer tissue was positively correlated with the expression level of HIF-1α （r=0.374， P<0.001） and the expression level of Sox2 （r=0.176， P<0.05）. The GEPIA Database analysis results showed that the cervical cancer patients with positive Gli1 expression had poor prognosis （P<0.05）. Under hypoxic status， the expression levels of Gli1， HIF-1α， and Sox2 mRNA and proteins in the SiHa cells were significantly increased （P<0.001）， and the cells exhibited a strong sphere-formation ability. Conclusion Hypoxia upregulates the expression levels of Gli1 and Sox2 mRNA and protein in the cervical cancer cells，and promotes the formation of stemness characteristics of the cancer cells.

Key words: Cervical neoplasm, Glioma-associated oncogene homolog 1, Hypoxia, Stemness, Hypoxia-inducible factor 1-alpha

CLC Number:

R737.33

Changji ZHU,Xingzhe LIU,Yanhua XUAN. Effect of hypoxia on expressions of Gli1 and Sox2 in cervical cancer SiHa cells[J].Journal of Jilin University(Medicine Edition), 2024, 50(2): 442-450.

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References 25

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Target gene	Sequence(5'－3')
HIF-1α	F:TTGCTCATCAGTTGCCACTTCC R:AGCAATTCATCTGTGCTTTCATGTC
Gli1	F:TCTGTGATGGGCAATGGTCT R.TCTGGGGTGGGATCAGGATA
Sox2	F:ATGCACCGCTACGACGTGA R:CTTTTGCACCCCTCCCATTT
Sox9	F-GAGGCCACGGAACAGACTCA R-CAGCGCCTTGAAGATAGCATT
CD44	F:CCAAGATGATCAGCCATTCTGG R.-AAGACATCTACCCCAGCAAC
OCT4	F-CAGAAGGGCAAGCGATCAAG R.-GGGCCAGAGGAAAGGACACT
LSD1	F-CTGTGCTTGTCCACCGAGTT R:ACCTGTTACCACCATGGCTC

Pathological diagnosis	n	Glil(－)	Glil(+)	χ²	P
Normal	41	28(68.3)	13(31.7)	5.322	0.150
CIN Ⅰ/Ⅱ	19	13(68.4)	6(31.6)
CIN Ⅲ	110	46(41.8)	64(58.2)
Squamous cell carcinoma	159	91(57.2)	68(42.8)

Variable	Total specimens(%)	Glil(－)	Glil(+)	χ²	P
Age（year）
<35	28(17.6)	15(53.6)	13(46.4)	0.301	0.583
≥35	131(82.4)	78(59.5)	53(40.5)	0.301	0.583
Lymphnode metastasis
Negative	121(76.1)	75(62.0)	46(38.0)	2.287	0.130
Positive	38(23.9)	34(89.5)	4(10.5)	2.287	0.130
Stage
Ⅰ	75( 47.2)	39(52.0)	36(48.0)	8.396	0.039
Ⅱ	22(13.8)	13(59.1)	9(40.9)
Ⅲ	34(21.4)	30(88.2)	4(11.8)
Ⅳ	28(17.6)	11(39.3)	17(60.7)

Variable	Total specimens	Glil(-)	Glil(+)	χ²	r	P
Sox2
Negative	104(65.4)	70(67.3)	34(32.7)	4.834	0.176	0.028
Positive	55(34.6)	16(29.1)	39(70.9)	4.834	0.176	0.028
Sox9
Negative	111(69.8)	63(56.8)	48(43.2)	1.244	0.091	0.265
Positive	48(30.2)	23(47.9)	25(52.1)	1.244	0.091	0.265
CD44
Negative	77(48.4)	42(54.5)	35(45.5)	0.705	0.063	0.401
Positive	82(51.6)	43(52.4)	39(47.6)	0.705	0.063	0.401
OCT4
Negative	95(59.7)	53(55.8)	42(44.2)	0.516	0.057	0.473
Positive	64(40.3)	32(50.0)	32(50.0)	0.516	0.057	0.473
LSD1
Negative	77(48.4)	37(48.1)	40(51.9)	1.498	0.128	0.221
Positive	82(51.6)	48(58.5)	34(41.5)	1.498	0.128	0.221

Variable	Total specimen	Glil(－)	Glil(+)	χ²	r	P
HIF-1α
Negative	67(42.1)	51(76.1)	16(23.9)	21.346	0.374	<0.001
Positive	92(57.9)	34(37.0)	58(63.0)	21.346	0.374	<0.001

Effect of hypoxia on expressions of Gli1 and Sox2 in cervical cancer SiHa cells

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Variable	Total specimens	HIF-1α(－)	HIF-1α(+)	χ²	r	P
Sox2
Negative	104(65.4)	56(53.8)	48(46.2)	12.020	0.294	0.001
Positive	55(34.6)	11(20.0)	44(80.0)	12.020	0.294	0.001
Sox9
Negative	111(69.8)	54(48.6)	57(51.4)	5.841	0.216	0.016
Positive	48(30.2)	13(27.1)	35(72.9)	5.841	0.216	0.016
CD44
Negative	77(48.4)	41(51.9)	36(48.1)	4.795	0.189	0.029
Positive	82(51.6)	26(31.7)	56(68.3)	4.795	0.189	0.029
OCT4
Negative	95(59.7)	49(51.6)	46(48.4)	7.811	0.248	0.005
Positive	64(40.3)	18(28.1)	46(71.9)	7.811	0.248	0.005