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Article Open Access

ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 in vitro

  • Authors:
    • Yihe Ma
    • Ye Chen
    • Yuhui Liang
    • Yun Huang
    • Miao Gong
    • Lifeng Tian
    • Xiaojun Xiao
    • Jingyan Liu
    • Qinmiao Huang
  • View Affiliations / Copyright

    Affiliations: Department of Respiratory Medicine and Allergy, The Third Affiliated Hospital (The Affiliated Luohu Hospital) of Shenzhen University, Shenzhen, Guangdong 518000, P.R. China, Department of Pediatric Medicine, Shenzhen Nanshan People's Hospital, Shenzhen, Guangdong 518000, P.R. China, Shenzhen Key Laboratory of Allergy and Immunology, State Key Laboratory of Respiratory Disease Shenzhen University Division, Shenzhen University Medical School, Shenzhen University, Shenzhen, Guangdong 518000, P.R. China, Emergency Department, The Second Affiliated Hospital, School of Medicine, The Chinese University of Hong Kong, Longgang District People's Hospital of Shenzhen, Shenzhen, Guangdong 518100, P.R. China
    Copyright: © Ma et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 77
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    Published online on: December 31, 2025
       https://doi.org/10.3892/mmr.2025.13787
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Abstract

Recurrence and metastasis are the leading causes of poor prognosis and death in lung cancer, and the mechanism of cancer metastasis has not yet been fully elucidated. As a gut-specific homeobox (HOX) transcription factor, intestine-specific HOX (ISX) is a proto-oncogene induced by the inflammatory factor IL-6. Notably, ISX overexpression can induce the epithelial-mesenchymal transition (EMT) response, and promotes tumor cell migration and invasion. In the present study, a lung cancer cell model with overexpression of ISX was established by infecting lung cancer cells with lentivirus. Reverse transcription-quantitative polymerase chain reaction was first used to verify the expression of the EMT-related gene induced by ISX overexpression. Furthermore, transcriptome sequencing and analysis showed that the overexpression of ISX induced significant changes in the gene expression profile of human lung cancer cells. In addition, type I collagen α1 chain (COL1A1), a highly expressed gene in various tumor tissues and cells, was shown to promote tumor cell migration and invasion, possibly by promoting EMT, and was significantly upregulated in human lung cancer cells overexpressing ISX. These results suggested that ISX may promote lung cancer migration and invasion by increasing the expression of COL1A1. In addition, four drugs that are currently used to treat lung cancer were screened. Of these, Iressa® (gefitinib) was revealed to significantly inhibit the viability, migration and invasion of lung cancer cells that stably overexpress ISX by downregulating the expression of COL1A1. In conclusion, these findings may help to prevent tumor metastasis and spread, and the potential molecular mechanism by which ISX promotes the development and migration of lung cancer was suggested. The current findings provide novel targets, and a scientific basis for the prevention and treatment of lung cancer, which may reduce costs for patients, their families and society.
View Figures

Figure 1

Overexpression of the oncogenic
factor ISX promotes the expression of EMT-related genes. (A) ISX
overexpression in A549 cells was mediated by Lv infection and
observed by GFP fluorescence intensity. Scale bar, 200 µm. (B) ISX
protein expression levels were verified by western blot analysis.
(C) ISX mRNA expression levels were verified by RT-qPCR detection.
(D) Genes related to EMT (TWIST, Slug, VIM and ZEB1) and epithelial
cells (E-cadherin) were detected by RT-qPCR. *P<0.05;
**P<0.01, ***P<0.001. EMT, epithelial-mesenchymal transition;
ISX, intestine-specific homeobox; Lv, lentivirus; MOI, multiplicity
of infection; RT-qPCR, reverse transcription-quantitative
polymerase chain reaction; VIM, vimentin; ZEB1, zinc finger E-box
binding homeobox 1.

Figure 2

Overexpression of ISX induces
significant changes in the gene expression profile of human lung
cancer cells. (A) Pearson's correlation analysis comparing the ISX
overexpression group with the control group. (B) Principal
component analysis comparing the ISX overexpression group with the
control group. (C) Venn diagram showing the number of genes shared
between the two groups (ISX overexpression and control). (D)
Heatmap showing gene expression in the ISX overexpression and
control groups. Con, control; ISX, intestine-specific homeobox; Lv,
lentivirus

Figure 3

Screening of DEGS in cells with ISX
overexpression, and enrichment analyses of biological functions and
signaling pathways. (A) Volcano plot showing the number of genes
with significant differences in expression levels between the ISX
overexpression group and the control group, as well as the specific
distribution of DEGs. (B) Heatmap showing the expression of DEGs
that were upregulated in the ISX overexpression groups. (C) KEGG
analysis showing the top 20 enriched pathways of the DEGs that were
upregulated in the ISX overexpression group compared with the
control group. Bubble size represents the number of genes, color
shade represents Padj, and the enrichment ratio on the y-axis
represents the number of genes/total number of genes. (D) GO term
enrichment analysis of the DEGs that were upregulated in the ISX
overexpression group compared with the control group. The numbers
on the columns indicate the number of enriched genes in that
category. BP, biological process; CC, cellular component; Con,
control; DEG, differentially expressed gene; GO, Gene Ontology;
ISX, intestine-specific homeobox; KEGG, Kyoto Encyclopedia of Genes
and Genomes; Lv, lentivirus; MF, molecular function.

Figure 4

ISX may promote lung cancer migration
and invasion by upregulating COL1A1. (A) Upregulated differentially
expressed genes in A549 cells with ISX overexpression were selected
to construct the PPI network. (B) Five core hub genes were
identified in the PPI network. (C) Column chart showing the
difference in COL1A1 expression in ISX-overexpressing A549 cells in
the transcriptome sequencing results. (D) mRNA expression levels of
COL1A1 were verified by reverse transcription-quantitative
polymerase chain reaction. *P<0.05. COL1A1, type I collagen α1
chain; ISX, intestine-specific homeobox; Lv, lentivirus; PPI,
protein-protein interaction.

Figure 5

Cell Counting Kit 8 screening to
identify mature tumor-targeting drugs that can inhibit the activity
of lung cancer cells stably overexpressing ISX. Effects of (A)
bevacizumab, the active ingredient of Avastin®, (B)
erlotinib, the active ingredient of Tarceva®, (C)
cetuximab, the active ingredient of Erbitux® lung cancer
drug and (D) gefitinib, the active ingredient of
Iressa®, on the survival of lung cancer cells not
overexpressing ISX. (E) Effect of gefitinib on the survival of
ISX-overexpressing lung cancer cells. *P<0.05; ****P<0.0001;
ns, not significant. ISX, intestine-specific homeobox.

Figure 6

Mature tumor-targeting drug
Iressa® (gefitinib) may inhibit the migration and
invasion of ISX-overexpressing human lung cancer cells by
regulating COL1A1. A549 cells infected with a GFP-labeled ISX
overexpression vector were treated with different concentrations of
gefitinib (0, 25, 50 µg/ml) for 48 h, and (A) cell migration (wound
healing; scale bar, 500 µm, and (B) invasion (Transwell; scale bar,
100 µm) assays were performed. (C) Reverse
transcription-quantitative polymerase chain reaction was used to
detect the effect of gefitinib (25 µg/ml, 48 h) on the mRNA
expression levels of COL1A1 in ISX-overexpressing A549 cells.
*P<0.05; **P<0.01; ****P<0.0001. COL1A1, type I collagen
α1 chain; ISX, intestine-specific homeobox; OD, optical
density.
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Copy and paste a formatted citation
Spandidos Publications style
Ma Y, Chen Y, Liang Y, Huang Y, Gong M, Tian L, Xiao X, Liu J and Huang Q: ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 <em>in vitro</em>. Mol Med Rep 33: 77, 2026.
APA
Ma, Y., Chen, Y., Liang, Y., Huang, Y., Gong, M., Tian, L. ... Huang, Q. (2026). ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 <em>in vitro</em>. Molecular Medicine Reports, 33, 77. https://doi.org/10.3892/mmr.2025.13787
MLA
Ma, Y., Chen, Y., Liang, Y., Huang, Y., Gong, M., Tian, L., Xiao, X., Liu, J., Huang, Q."ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 <em>in vitro</em>". Molecular Medicine Reports 33.2 (2026): 77.
Chicago
Ma, Y., Chen, Y., Liang, Y., Huang, Y., Gong, M., Tian, L., Xiao, X., Liu, J., Huang, Q."ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 <em>in vitro</em>". Molecular Medicine Reports 33, no. 2 (2026): 77. https://doi.org/10.3892/mmr.2025.13787
Copy and paste a formatted citation
x
Spandidos Publications style
Ma Y, Chen Y, Liang Y, Huang Y, Gong M, Tian L, Xiao X, Liu J and Huang Q: ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 <em>in vitro</em>. Mol Med Rep 33: 77, 2026.
APA
Ma, Y., Chen, Y., Liang, Y., Huang, Y., Gong, M., Tian, L. ... Huang, Q. (2026). ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 <em>in vitro</em>. Molecular Medicine Reports, 33, 77. https://doi.org/10.3892/mmr.2025.13787
MLA
Ma, Y., Chen, Y., Liang, Y., Huang, Y., Gong, M., Tian, L., Xiao, X., Liu, J., Huang, Q."ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 <em>in vitro</em>". Molecular Medicine Reports 33.2 (2026): 77.
Chicago
Ma, Y., Chen, Y., Liang, Y., Huang, Y., Gong, M., Tian, L., Xiao, X., Liu, J., Huang, Q."ISX promotes tumor migration and invasion in lung cancer by upregulating COL1A1 <em>in vitro</em>". Molecular Medicine Reports 33, no. 2 (2026): 77. https://doi.org/10.3892/mmr.2025.13787
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