通用转录因子2I在胶质母细胞瘤替莫唑胺化疗抵抗中的作用

doi:10.13481/j.1671-587X.20240219

Abstract

Abstract:

Objective To discuss the effect of general transcription factor 2I （GTF2I） on the chemotherapeutic resistance to temozolomide in the glioblastoma multiforme （GBM）， and to clarify its mechanism. Methods Bioinformatics analysis was used to identify the common transcription factors for DNA methyltransferase 1 （DNMT1）， damage-specific DNA binding protein 1 （DDB1）， chromobox protein homolog 5 （CBX5）， and xeroderma pigmentosum complementation group C （XPC） in GBM tissue using the transcription factor prediction website （PROMO website）； correlation analysis and survival analysis for DDB1， CBX5， XPC， and DNMT1 with GTF2I and methylguanine methyltransferase （MGMT） were conducted based on The Cancer Genome Atlas （TCGA） Database；small interfering RNAs （siRNAs） were used to transfect and silence the gene expression of MGMT and GTF2I in the human T98 GBM cells and LN229 glioma cells； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of the gene mRNA. After silencing the GTF2I gene， plate clone formation assay was used to detect the colon forming ability of the tumor cells， and CCK-8 assay was used to detect the sensitivity of the cells to temozolomide. Results The bioinformatics analysis results showed that the expression levels of DDB1， CBX5， XPC， and DNMT1 were significantly positively correlated with the expression level of GTF2I in GBM tissue （P<0.05）， and were negatively correlated with the expression level of MGMT （P<0.05）. The expression level of GTF2I was significantly negatively correlated with the expression level of MGMT （P<0.05）. Excluding the GBM patients who did not receive temozolomide treatment， the survival analysis results indicated that the patients with high expression of GTF2I had a decreased overall survival time. After silencing the MGMT gene， the expression levels of GTF2I， DDB1， CBX5， and XPC mRNA in the human brain glioma T98 cells were increased （P<0.001）； after silencing the GTF2I gene， the expression level of MGMT mRNA in the human brain glioma LN229 cells was increased （P<0.05）， while the expression levels of DDB1， CBX5， XPC， and DNMT1 mRNA were significantly decreased （P<0.05 or P<0.001）. The plate clone formation assay results showed that there was no significant difference in the clone formation ability of the cells before and after silencing the GTF2I gene （P=0.138）. The CCK-8 assay results showed that compared with control group，the viability of the cells in observation group was decreased （P<0.05）. Conclusion The transcription factor GTF2I regulates the expressions of key DNA damage repair protein mRNA， including MGMT， DDB1， CBX5， and XPC， and is involved in the chemotherapeutic resistance of the GBM cells to temozolomide；GTF2I may represent a potential new therapeutic target for the GBM.

Key words: Glioblastoma multiforme, Temozolomide resistance, Universal transcription factor 2I, Methylguanine methyltransferase, Key DNA damage repair genes

CLC Number:

R739.41

Jianguo ZHOU,Hongjian JIANG,Qihui ZHU,Gengqiang ZHANG,Qilin DENG,Ling QI,Kaishu LI,Hongquan YU. Effect of general transcription factor 2I on temozolomide chemotherapy resistance of glioblastoma[J].Journal of Jilin University(Medicine Edition), 2024, 50(2): 457-464.

Figures/Tables 7

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References 23

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Primer	Sequence (5′－3′)
GTF2I-1	Forward Reverse	GGCCAAGUCCAAAGCCGAATT UUCGGCUUUGGACUUGGCCTT
GTF2I-2	Forward Reverse	GGUGGUCGUGUGAUGGUAATT UUACCAUCACACGACCACCTT
GTF2I-3	Forward Reverse	CCACAGAAGAUUCUGGCAUTT AUGCCAGAAUCUUCUGUGGTI
MGMT-1	Forward Reverse	GCAGGGUCUGCACGAAAUATT UAUUUCGUGCAGACCCUGCTT
MGMT-2	Forward Reverse	GCAAUGAGAGGCAAUCCUGTT CAGGAUUGCCUCUCAUUGCTT
MGMT-3	Forward Reverse	GCUGCUGAAGGUUGUGAAATT UUUCACAACCUUCAGCAGCTT

Primer	Sequence (5′－3′)
GTF2I	Forward Reverse	GCATGTAGGTGGTCGTGTGA CAGTTTCAGAAGGGCCTTGA
MGMT	Forward Reverse	GAAGCTGGAGCTGTCTGGTT CGGGGAACTCTTCGATAGCC
CBX5	Forward Reverse	TAGACAGGCGCGTGGTTAAG AGCCCCGAGCGATATCATTG
DDB1	Forward Reverse	CAAAGCACGATTGTGTGCCA TGAGCTTCACAAGCTGGGAG
XPC	Forward Reverse	TGCCTATTGGCTGTGTCCAG AGACCTTTGGCCAGCAACTT
DNMT1	Forward Reverse	AGGAGGGCTACCTGGCTAAA GCTTAGCCTCTCCATCGGAC

Effect of general transcription factor 2I on temozolomide chemotherapy resistance of glioblastoma

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