HOXD10 Expression Research Articles

RETRACTION: MeCP2 Drives Hepatocellular Carcinoma Progression Via Enforcing HOXD3 Promoter Methylation and Expression Through the HB-EGF/EGFR Pathway.

L. Wang, Y. Gao, D. Tong, X. Wang, C. Guo, B. Guo, Y. Yang, L. Zhao, J. Zhang, J. Yang, Y. Qin, L. Liu, and C. Huang, "MeCP2 Drives Hepatocellular Carcinoma Progression Via Enforcing HOXD3 Promoter Methylation and Expression Through the HB-EGF/EGFR Pathway," Molecular Oncology 15, no. 11 (2021): 3147-3163, https://doi.org/10.1002/1878-0261.13019. The above article, published online on 24 May 2021 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal Editor-in-Chief, Kevin Ryan; FEBS Press; and John Wiley and Sons Ltd. The retraction has been agreed upon following an investigation into concerns raised by a third party regarding an image duplication between Figures 2C (MeCP2 panel) and 4G (left panel). The investigation also revealed inconsistencies in the gel blots of Figures 2J and 3N. The partial raw data shared by the authors was deemed insufficient to address these concerns. Therefore, the editors have lost confidence in the presented data, consider the conclusions substantially compromised and are retracting the paper. The authors were informed of the decision to retract but did not agree to the retraction or the retraction wording.

Adaptive tail-length evolution in deer mice is associated with differential Hoxd13 expression in early development

Variation in the size and number of axial segments underlies much of the diversity in animal body plans. Here we investigate the evolutionary, genetic and developmental mechanisms driving tail-length differences between forest and prairie ecotypes of deer mice (Peromyscus maniculatus). We first show that long-tailed forest mice perform better in an arboreal locomotion assay, consistent with tails being important for balance during climbing. We then identify six genomic regions that contribute to differences in tail length, three of which associate with caudal vertebra length and the other three with vertebra number. For all six loci, the forest allele increases tail length, indicative of the cumulative effect of natural selection. Two of the genomic regions associated with variation in vertebra number contain Hox gene clusters. Of those, we find an allele-specific decrease in Hoxd13 expression in the embryonic tail bud of long-tailed forest mice, consistent with its role in axial elongation. Additionally, we find that forest embryos have more presomitic mesoderm than prairie embryos and that this correlates with an increase in the number of neuromesodermal progenitors, which are modulated by Hox13 paralogues. Together, these results suggest a role for Hoxd13 in the development of natural variation in adaptive morphology on a microevolutionary timescale.

Inhibition of HOXD11 promotes cartilage degradation and induces osteoarthritis development

The 5′-HOXD genes are important for chondrogenesis in vertebrates, but their roles in osteoarthritis (OA) are still ambiguous. In our study, 5′-HOXD genes involvement contributing to cartilage degradation and OA was investigated. In bioinformatics analysis of 5′-HOXD genes, we obtained the GSE169077 data set related to OA in the GEO and analyzed DEGs using the GEO2R tool attached to the GEO. Then, we screened the mRNA levels of 5′-HOXD genes by quantitative reverse transcriptase–polymerase chain reaction (qRT-PCR). We discovered that OA chondrocyte proliferation was inhibited, and apoptosis was increased. Moreover, it was discovered that SOX9 and COL2A1 were downregulated at mRNA and protein levels, while matrix metalloproteinases (MMPs) and a disintegrin-like and metalloproteinase with thrombospondin motifs (ADAMTSs) were upregulated. According to the results of differentially expressed genes (DEGs) and qRT-PCR, we evaluated the protein level of HOXD11 and found that the expression of HOXD11 was downregulated, reversed to MMPs and ADAMTSs but consistent with the cartilage-specific factors, SOX9 and COL2A1. In the lentivirus transfection experiments, HOXD11 overexpression reversed the effects in OA chondrocytes. In human OA articular cartilage, aberrant subchondral bone was formed in hematoxylin–eosin (H&E) and Safranin O and fast green (SOFG) staining results. Furthermore, according to immunohistochemistry findings, SOX9 and HOXD11 expression was inhibited. The results of this study established that HOXD11 was downregulated in OA cartilage and that overexpression of HOXD11 could prevent cartilage degradation in OA.

Ropivacaine combined with sorafenib attenuates hepatocellular carcinoma cell proliferation and metastasis by inhibiting the miR-224/HOXD10 axis.

The development of drug resistance in hepatocellular carcinoma (HCC) cells limits the effectiveness of sorafenib (Sor). However, the regulatory mechanisms underlying the effects of the combination Sor and ropivacaine (Rop) on HCC cells remain unclear. miR-224 and HOXD10 mRNA expression in HCC cells was analyzed using qRT-PCR. CCK-8, Transwell assays and tumor formation experiments in nude mice were used to assess HCC cell proliferation, migration, and invasion. Migration of HCC cells was also analyzed using a cell scratch assay. Hematoxylin and eosin staining was used to detect tumor area. miR-224 expression profoundly increased in HepG2 and Huh7 cells. Treatment with Rop and/or Sor blocked miR-244 expression, especially the combination treatment. Transfection of miR-224 mimic increased HCC cell proliferation and tumor size in nude mice, and migration and invasion in vitro in the presence of Rop and Sor compared to the negative control mimic. Dual-luciferase reporter assays showed that HOXD10 was targeted by miR-224. HOXD10 protein expression and was markedly reduced in HepG2 and Huh7 cells. Rop and/or Sor treatment increased HOXD10 protein expression, particularly the combination treatment. miR-224 negatively regulated HOXD10 expression in HCC cells treated with Rop and Sor. Transfection-mediated silencing of HOXD10 increased HCC cell proliferation, migration, and invasion in the presence of Rop and Sor compared with negative control transfection. The combination of Rop and Sor attenuates HCC cell proliferation and metastasis via the miR-224/HOXD10 axis.

HOXD9 is a potential prognostic biomarker involved in immune microenvironment of glioma.

Glioma is the prevailing malignant tumor affecting the brain and central nervous system, constituting over 80% of all malignant brain tumors. HOXD9 has been implicated in the development of glioma, but the specific mechanism of its influence on glioma pathogenesis remains incompletely understood. The purpose of this study was to investigate the role of HOXD9 in glioma and examine the changes in HOXD9 expression during the progression of glioma, thus contributing new insights into the pathogenesis of glioma. Glioma samples from the Cancer Gene Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) datasets were included in this study. Variations in HOXD9 expression in gliomas between different subgroups of multiple clinical characteristics were explored, and the expression was validated in glioma samples using qRT-PCR and western blotting. Next, the impact of HOXD9 on the prognosis of gliomas was explored by survival analysis, receiver operating characteristic curve, and nomogram plots. Subsequently, the association between HOXD9 and the tumor immune microenvironment was explored using the ssGSEA algorithm and the ESTIMATE algorithm. Then, immune-related pathways associated with HOXD9 were determined by differential express analysis and GSEA. Finally, HOXD9-related genomic alterations were identified. HOXD9 expression is upregulated and correlated with malignant properties in glioma. Similarly, our validation results showed significantly upregulated protein and mRNA levels of HOXD9 in glioma brain tissues. In addition, high HOXD9 expression was indicative of a poor prognosis for glioma patients. Additionally, elevated HOXD9 levels were associated with reduced tumor purity and higher levels of immune invasion. Finally, HOXD9 was significantly associated with genomic alterations. Overall, this study has unveiled a significant association between HOXD9 and the prognosis and survival of glioma patients. Our findings highlight the potential of HOXD9 as a prognostic biomarker, implicating its role in influencing the glioma immune microenvironment.

Homeobox D9 Promotes Malignant Progression of Non-Small Cell Lung Cancer Through Negatively Regulating Kallikrein Related Peptidase 9

We investigated the association between HOXD9 expression and prognosis in non-small cell lung cancer (NSCLC) and explored its underlying mechanisms in tumor progression. HOXD9 expression was examined in 42 pairs of NSCLC tissue samples using qRT-PCR, and its effects were evaluated in NSCLC cell lines through lentiviral vector transfection, assessing cell growth and migration using CCK-8 and Transwell assays. Bioinformatics analysis identified KLK9 as a target gene of HOXD9, and their interaction was investigated in vitro and in vivo. HOXD9 mRNA expression was significantly higher in NSCLC tissues compared to adjacent tissues, positively correlating with metastasis incidence and poor prognosis. HOXD9 knockdown reduced NSCLC cell migration and proliferation, while overexpression had the opposite effect. Luciferase assays confirmed HOXD9′s binding to KLK9, leading to negative regulation of KLK9 expression. Knockdown of KLK9 reversed the inhibitory effect of HOXD9 on NSCLC cell migration and proliferation, while co-overexpression of KLK9 and HOXD9 had the opposite effect. Our findings demonstrate the close association of high HOXD9 expression in NSCLC tissues with metastasis incidence and poor prognosis. HOXD9 likely exerts its biological function in NSCLC by negatively regulating KLK9. These results provide valuable insights into the molecular mechanisms driving NSCLC progression, highlighting HOXD9 as a potential prognostic biomarker and therapeutic target.

The shh limb enhancer is activated in patterned limb regeneration but not in hypomorphic limb regeneration in Xenopus laevis

Xenopus young tadpoles regenerate a limb with the anteroposterior (AP) pattern, but metamorphosed froglets regenerate a hypomorphic limb after amputation. The key gene for AP patterning, shh, is expressed in a regenerating limb of the tadpole but not in that of the froglet. Genomic DNA in the shh limb-specific enhancer, MFCS1 (ZRS), is hypermethylated in froglets but hypomethylated in tadpoles: shh expression may be controlled by epigenetic regulation of MFCS1. Is MFCS1 specifically activated for regenerating the AP-patterned limb? We generated transgenic Xenopus laevis lines that visualize the MFCS1 enhancer activity with a GFP reporter. The transgenic tadpoles showed GFP expression in hoxd13-and shh-expressing domains of developing and regenerating limbs, whereas the froglets showed no GFP expression in the regenerating limbs despite having hoxd13 expression. Genome sequence analysis and co-transfection assays using cultured cells revealed that Hoxd13 can activate Xenopus MFCS1. These results suggest that MFCS1 activation correlates with regeneration of AP-patterned limbs and that re-activation of epigenetically inactivated MFCS1 would be crucial to confer the ability to non-regenerative animals for regenerating a properly patterned limb.

Abstract P3-08-02: Prospective Use of Feline Mammary Carcinomas to Study MiR-10b and Metastasis

Abstract Despite many advances in cancer treatment, metastatic disease is estimated to be responsible for 90% of all cancer-related deaths. Current treatments for metastatic disease target various aspects of carcinogenesis but not specifically the metastatic process, representing a major unmet clinical need. MicroRNA-10b, a small non-coding RNA, offers tremendous potential as a treatment target for metastatic disease. It is implicated in invasion, migration, and viability of metastatic cells across a variety of cancer types, and it is upregulated in metastases compared to their matched primary tumor, establishing miR-10b as a potential treatment target unique to metastatic niche. We have previously developed a therapeutic targeting miR-10b in metastases and tested it in murine models of metastatic breast cancer. This therapeutic, consisting of an anti-miR-10b antisense oligonucleotide conjugated to iron oxide-based nanoparticles, prevents metastasis and eradicates pre-existing metastases in murine breast cancer models. With an outlook to clinical translation of our approach, we seek to test our therapeutic strategy in larger animal models. Feline mammary carcinomas are considered by most to be the best large animal model for human breast cancer due to similarities such as relative age of onset, histopathology, metastatic patterns, and treatment response. To support the use of this model with our therapeutic, we investigated the characteristics of miR-10b expression in spontaneous metastatic breast cancer in companion cats. Archival blocks of matched primary tumors and metastatic lymph nodes from companion cats diagnosed with mammary carcinoma (n=9, 44%TNBC, 56%HER2+) were obtained from the tissue bank of the Michigan State University (MSU) Veterinary Diagnostic Laboratory (VDL). Tissues were analyzed for miR-10b and its target HOXD10 expression using qRT-PCR and in situ hybridization. qRT-PCR revealed that miR-10b expression was significantly upregulated in 55.5% of lymph node metastases compared to their matched primary tumor, mirroring findings in human metastatic cancer. This was validated by qRT-PCR for HOXD10 gene expression (a direct target of miR-10b), which was significantly downregulated in these metastases compared to their matched primary tumor. In situ hybridization demonstrated that miR-10b expression was increased at the invasive edge of tumors and in actively invading cells, suggesting miR-10b plays a similar role in invasion in feline breast cancer as it does in human breast cancer. Altogether, these findings support the use of feline mammary carcinomas as a model of human breast cancer and as an excellent candidate for treatment with our therapeutic. Citation Format: Alan Halim, N. Anna Savan, Paulo Vilar Saavedra, Vilma Yuzbasiyan-Gurkan, Matti Kiupel, Lorenzo Sempere, Anna Moore. Prospective Use of Feline Mammary Carcinomas to Study MiR-10b and Metastasis [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P3-08-02.

A novel Hoxd13 mutation causes synpolydactyly and promotes osteoclast differentiation by regulating pSmad5/p65/c-Fos/Rank axis

The mutations of HOXD13 gene have been involved in synpolydactyly (SPD), and the polyalanine extension mutation of Hoxd13 gene could lead to SPD in mice. In this study, a novel missense mutation of Hoxd13 (NM_000523: exon2: c.G917T: p.R306L) was identified in a Chinese family with SPD. The mice carrying the corresponding Hoxd13mutation were generated. The results showed that the homozygous mutation of Hoxd13 also caused SPD, but heterozygous mutation did not affect limbs development, which was different from that of SPD patients. With the increasing generation, the mice with homozygous Hoxd13 mutation presented more severe syndactyly. Western blotting showed that this mutation did not affect the protein expression of Hoxd13, suggesting that this mutation did not result in haploinsufficiency. Further analysis demonstrated that this homozygous Hoxd13mutation promoted osteoclast differentiation and bone loss, and enhanced the mRNA and protein expression of osteoclast-related genes Rank, c-Fos, and p65. Meanwhile, this homozygous Hoxd13 mutation elevated the level of phosphorylated Smad5 (pSmad5). Co-immunoprecipitation verified that this mutation attenuated the interaction between pSmad5 and HOXD13, suggesting that this mutation released more pSmad5. Inhibition of pSmad5 reduced the expression of Rank, c-Fos, and p65 despite in the mutation group. In addition, inhibition of pSmad5 repressed the osteoclast differentiation. ChIP assay confirmed that p65 and c-Fos could bind to the promoter of Rank. These results suggested that this novel Hoxd13 mutation promoted osteoclast differentiation by regulating Smad5/p65/c-Fos/Rank axis, which might provide a new insight into SPD development.

Long Noncoding RNA LINC00909 Induces Epithelial-Mesenchymal Transition and Contributes to Osteosarcoma Tumorigenesis and Metastasis.

Background Osteosarcoma (OS) is a malignant tumor that is highly metastatic with a high mortality rate. Although mounting evidence suggests that LINC00909 is strongly associated with the malignant progression of various tumors, the exact role of LINC00909 in OS remains unknown. Therefore, the current study was designed to investigate the relationship between LINC00909 and the malignant progression of OS. Methods LINC00909 expression was measured in OS cell lines and clinical specimens using RT-qPCR assays. The effects of LINC00909 on OS proliferation, invasion, and migration were calculated both in vitro and in vivo. Apart from that, bioinformatics analyses, FISH, RIP, and luciferase reporter assays were carried out to investigate the downstream target of LINC00909. Rescue experiments were also conducted to investigate the potential mechanisms of action of competitive endogenous RNAs (ceRNAs). Results In this study, we found that LINC00909 was highly expressed in OS cell lines and clinical specimens. In vivo and in vitro experiments demonstrated that LINC00909 induces epithelial-to-mesenchymal transition (EMT) and contributes to OS tumorigenesis and metastasis. FISH, RIP, and luciferase assays indicated that miR-875-5p is a direct target of LINC00909. Moreover, HOXD9 was validated as the downstream target of miR-875-5p in a luciferase reporter assay and western blotting experiments. Rescue experiments revealed that HOXD9 reversed the effect of LV-sh-LINC00909 on OS cells by positively regulating the PI3K/AKT/mTOR signaling pathway. Conclusion Collectively, LINC00909 induces EMT and contributes to OS tumorigenesis and metastasis through the PI3K/AKT/mTOR pathway by binding to miR-875-5p to upregulate HOXD9 expression. Targeting the LINC00909/miR-875-5p/HOXD9 axis may have potential in treating OS.

Molecular Analysis of Prognosis and Immune Infiltration of Ovarian Cancer Based on Homeobox D Genes.

Background Homeobox D (HOXD) genes were associated with cancer pathogenesis. However, the role of HOXD genes in ovarian cancer (OC) and the possible mechanisms involved are unclear. In this study, we analyzed the function and regulatory mechanisms and functions of HOXD genes in OC based on comprehensive bioinformatics analysis. Methods Expression of HOXD1/3/4/8/9/10/11/12/13 mRNA was analyzed between OC tissue and normal tissue using ONCOMINE, GEO, and TCGA databases. The relationship between HOXD expression and clinical stage was studied by GEPIA. The Kaplan-Meier plotter was used to analyze prognosis. cBioPortal was used to analyze the mutation and coexpression of HOXDs. GO and KEGG analyses were performed by the DAVID software to predict the function of HOXD coexpression genes. Immune infiltration analysis was used to evaluate the relationship between the expression of HOXD genes and 24 immune infiltrating cells. Results The expression of HOXD3/4/8/9/10/11 was significantly lower in OC tissues than in normal ovarian tissues, while the expression of HOXD1/12/13 was significantly higher in OC tissues. The expression of HOXD genes was associated with FIGO stage, primary therapy outcome, tumor status, anatomic neoplasm subdivision, and age. The expression levels of HOXD1/3/4/8/9/10 correlated with tumor stage. HOXD1/8/9 could be served as ideal biomarkers to distinguish OC from normal tissue. Low HOXD9 expression was associated with shorter overall survival (OS) (HR: 0.75; 95% CI: 0.58–0.98; P = 0.034) and progression-free survival (PFS) (HR: 0.69; 95% CI: 0.54–0.87; P = 0.002). The HOXD coexpression genes were associated with pathways including cell cycle, TGF-beta signaling pathway, cellular senescence, and Hippo signaling pathway. HOXD genes were significantly associated with immune infiltration. Conclusion The expression of HOXD genes is associated with clinical characteristics. HOXD9 is a new biomarker of prognosis in OC, and HOXD1/4/8/9/10 may be potential therapeutic targets. The members of the HOXD genes may be the response to immunotherapy for OC.

MiR-138-5p-mediated HOXD11 promotes cell invasion and metastasis by activating the FN1/MMP2/MMP9 pathway and predicts poor prognosis in penile squamous cell carcinoma

The presence and extent of regional lymph node and distant metastasis are the most fatal prognostic factors in penile squamous cell carcinoma (PSCC). However, the available biomarkers and detailed mechanisms underlying the metastasis of PSCC remain elusive. Here, we explored the expression landscape of HOX genes in twelve paired PSCC tissues, including primary tumors, metastatic lymph nodes and corresponding normal tissues, and highlighted that HOXD11 was indispensable in the progression of PSCC. HOXD11 was upregulated in PSCC cell lines and tumors, especially in metastatic lymph nodes. High HOXD11 expression was associated with aggressive features, such as advanced pN stages, extranodal extension, pelvic lymph node and distant metastasis, and predicted poor survival. Furthermore, tumorigenesis assays demonstrated that knockdown of HOXD11 not only inhibited the capability of cell proliferation, invasion and tumor growth but also reduced the burden of metastatic lymph nodes. Further mechanistic studies indicated that miR-138-5p was a tumor suppressor in PSCC by inhibiting the translation of HOXD11 post-transcriptionally through binding to the 3′ untranslated region. Furthermore, HOXD11 activated the transcription of FN1 to decompose the extracellular matrix and to promote epithelial mesenchymal transition-like phenotype metastasis via FN1/MMP2/MMP9 pathways. Our study revealed that HOXD11 is a promising prognostic biomarker and predicts advanced disease with poor outcomes, which could serve as a potential therapeutic target for PSCC.

A Druggable Rheostat for Ewing Sarcoma?

The posterior HOXD enhancer is an EWSR1::FLI1-dependent regulator of HOXD13 expression in Ewing sarcoma. HOXD13 expression promotes a mesenchymal cell state. Through antagonistic transcriptional programs, EWSR1::FLI1 and HOXD13 serve as master regulators of Ewing cell plasticity. Targeting Ewing cells as they exist in/transition between mesenchymal states is a priority. See related article by Apfelbaum et al., p. 4466.

Effects of toxic Microcystis aeruginosa on the expression of Hox genes in Daphnia similoides sinensis.

Lake eutrophication and cyanobacterial blooms have become worldwide environmental issues. Under cyanobacterial blooms (especially Microcystis), Daphnia spp. can transfer beneficial information to their offspring in order to improve adaptability. Hox genes are important regulatory factors of transcription in metazoans, and are involved in the growth and development of organisms. However, the mechanisms of Microcystis on the expression of Hox genes in Daphnia are unclear. In this study, the effects of Microcystis aeruginosa on Hox gene expression in the mothers and offspring (F1) of two Daphnia similoides sinensis clones were investigated using a mixed diet of M. aeruginosa and Scenedesmus obliquus. Compared with the 100%S food treatment, the survival rates at the end of the experiment of clone 1‐F1 in the food treatments containing M. aeruginosa were significantly lower, but it was significantly higher for clone 2‐F1 in the 20%M + 80%S food treatment. Moreover, the survival rates at the end of the experiment of clone 1‐F1 in the food treatments containing M. aeruginosa were significantly higher than those of their mother. Based on previous transcriptome data, 14 Hox genes of D. similoides sinensis were identified, including Abd‐B, CDX‐1, Dll, HOX‐1, HOX‐2, HOXA1, HOXA2, HOXB3, HOXB3‐2, HOXB7, HOXC4, HOXC7, HOXC8, and HOXD10. The expressions of Abd‐B, HOX‐2, HOXA1, HOXC7, and HOXD10 of clone 2‐mothers in the 40%M + 60%S food treatment were 2.9–22.5 times as high as in the 100%S food treatment, whereas the expressions of CDX‐1, HOX‐1, HOXB3, and HOXD10 of clone 1‐mothers were 4.8–13.1 times at same food level. The expression of HOXA2, HOXC7, HOXC8, and HOXD10 of clone 1‐F1 in the 40%M + 60%S food treatment was 8.2–21.1 times as high as in the 100%S food treatment. However, compared with the 100%S food treatment, the expressions of CDX‐1 in the mothers and F1 of clone 2 and HOXB7 in the mothers of clone 1 in the food treatments containing M. aeruginosa were significantly lower (p < .05). Our results suggest that the offspring (F1) produced by D. similoides sinensis mother pre‐exposed to toxic M. aeruginosa had stronger adaptability to M. aeruginosa than their mothers. Moreover, Hox gene expressions of D. similoides sinensis had obvious differences between clones under stress of toxic M. aeruginosa.

Newt Hoxa13 has an essential and predominant role in digit formation during development and regeneration.

The 5'Hox genes play crucial roles in limb development and specify regions in the proximal-distal axis of limbs. However, there is no direct genetic evidence that Hox genes are essential for limb development in non-mammalian tetrapods or for limb regeneration. Here, we produced single to quadruple Hox13 paralog mutants using the CRISPR/Cas9 system in newts (Pleurodeles waltl), which have strong regenerative capacities, and also produced germline mutants. We show that Hox13 genes are essential for digit formation in development, as in mice. In addition, Hoxa13 has a predominant role in digit formation, unlike in mice. The predominance is probably due to the restricted expression pattern of Hoxd13 in limb buds and the strong dependence of Hoxd13 expression on Hoxa13. Finally, we demonstrate that Hox13 genes are also necessary for digit formation in limb regeneration. Our findings reveal that the general function of Hox13 genes is conserved between limb development and regeneration, and across taxa. The predominance of Hoxa13 function both in newt limbs and fish fins, but not in mouse limbs, suggests a potential contribution of Hoxa13 function in fin-to-limb transition.

Developmental biology: A 5ʹHoxd–Gli3 balance in tetrapod axial polarity

Almost all living tetrapods exhibit postaxial dominance in digit formation, apart from urodele amphibians, which show preaxial dominance. Recent work shines light on the genetic differences between the two modes of limb development, suggesting that differences in 5'Hoxd expression, mediated by Gli3, may explain the switch in axial polarity.

Restoring HOXD10 Exhibits Therapeutic Potential for Ameliorating Malignant Progression and 5-Fluorouracil Resistance in Colorectal Cancer.

Emerging evidence suggests that hypermethylation of HOXD10 plays an important role in human cancers. However, the biological and clinical impacts of HOXD10 overmethylation and its downstream targets in colorectal cancer remain unknown. We evaluated the methylation level of HOXD10 in paired cancer and normal tissues (n = 42) by using pyrosequencing, followed by validation of the methylation status of HOXD10 from The Cancer Genome Atlas (TCGA) datasets with 302 cancer tissues and 38 normal tissues. The biological function of HOXD10 was characterized in cell lines. We further evaluated the effects of HOXD10 and its targets on chemoresistance in our established resistant cell lines and clinical cohort (n = 66). HOXD10 was found frequently methylated in colorectal cancer, and its hypermethylation correlates with its low expression level, advanced disease, and lymph node metastasis. Functionally, HOXD10 acts as a tumor suppressor gene, in which HOXD10-expressing cells showed suppressed cell proliferation, colony formation ability, and migration and invasion capacity. Mechanistically, DNMT1, DNMT3B, and MeCP2 were recruited in the HOXD10 promoter, and demethylation by 5-Aza-2′-deoxycytidine (5-Aza-CdR) treatment or MeCP2 knockdown can sufficiently induce HOXD10 expression. HOXD10 regulates the expressions of miR-7 and IGFBP3 in a promoter-dependent manner. Restoration of the expression of HOXD10 in 5-fluorouracil (5-FU)-resistant cells significantly upregulates the expressions of miR-7 and IGFBP3 and enhances chemosensitivity to 5-FU. In conclusion, we provide novel evidence that HOXD10 is frequently methylated, silenced, and contributes to the development of colorectal cancers. Restoration of HOXD10 activates the expressions of miR-7 and IGFBP3 and results in an inhibited phenotype biologically, suggesting its potential therapeutic relevance in colorectal cancer (CRC).

Identification of HOXD10 as a Marker of Poor Prognosis in Glioblastoma Multiforme.

PurposeHOXD10 is a tumor modulator that can either be a tumor-suppressor or a tumor-promoting gene. However, the role of HOXD10 in glioblastoma multiforme (GBM) remains unclear.MethodsImmunohistochemistry (IHC) was applied to detect protein expression of HOXD10 in GBM and normal brain tissue patients. Clinicopathological characteristics with GBM were recorded, and a Kaplan–Meier curve was plotted. Additionally, the mRNA expression of HOXD10 and its effect on prognosis were analyzed using the online tool GEPIA and the Cancer Genome Atlas (TCGA), Chinese Glioma Genome Atlas (CGGA), and the Gene Expression Omnibus (GEO) databases. Based on the mRNA expression of HOXD10, GBM patients from TCGA database were divided into low- and high-HOXD10 expression groups to analyze the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and construct a lncRNA-miRNA-mRNA network and a protein–protein interaction (PPI) network.ResultsThe mRNA expression of HOXD10 was up-regulated in GBM according to GEPIA, while the protein expression of HOXD10 in GBM was down-regulated according to IHC analysis of samples from patients collected from our hospital. Correlation analysis showed that HOXD10 expression was significantly related to IDH1 status. Univariate analysis revealed that low HOXD10 expression, complete surgical resection, postoperative radiotherapy, postoperative temozolomide chemotherapy and IDH1 mutation were all beneficial prognostic factors. Further multivariate analysis revealed that only complete surgical resection and postoperative radiotherapy were independent prognostic factors. GO and KEGG enrichment analyses indicated that HOXD10 expression is mainly involved in cytokine-cytokine receptor interactions. In the ceRNA network, 89 nodes, containing 45 mRNAs, 39 miRNAs and five lncRNAs associated with prognosis were involved. The PPI network revealed a tight interaction between HOXD10 and HOXD8, HOXD9, HOXD11, HOXD13 and HOXB3.ConclusionBased on our experimental data, although HOXD10 expression is low in GBM compared with normal brain tissue, GBM patients with high HOXD10 expression have a worse prognosis. HOXD10 may play different or even opposite roles in different stages of GBM occurrence and development. For patients with GBM, HOXD10 may be a valid predictor of prognosis.

HOXD13 promotes the malignant progression of colon cancer by upregulating PTPRN2.

PurposeThe homeobox (HOX) family plays an important role in multi‐biological processes, such as morphogenesis and tumors. However, the function of HOXD13 in colon cancer remains unclear.Materials and MethodsThe Cancer Genome Atlas database was used to analyze the expression of HOXD13 and its effect on the survival rate of colon cancer patients. Wound healing, Transwell, and clone formation were used to evaluate the effects of changes in HOXD13 expression on the function of colon cancer cells. A nude mouse xenograft tumor model was used to test the effects of HOXD13 on tumor growth in vivo.ResultsOur results showed that HOXD13 was highly expressed in colon cancer and predicted a poor prognosis for patients. In in vitro experiments, the knockdown of HOXD13 can inhibit the proliferation and invasion of colon cancer cells. In vivo experiments showed the inhibited tumor growth after the knockdown of HODX13. In addition, HOXD13 bound to the protein tyrosine phosphatase receptor type N2 (PTPRN2) promoter and promoted the transcription of PTPRN2.ConclusionWe revealed the function and mechanism of HOXD13 in colon cancer and suggest that HOXD13 may be a candidate marker for the diagnosis and treatment of colon cancer.

Prognostic and clinical significance of HOXC9 and HOXD10 in papillary thyroid cancer.

BackgroundThe homebox superfamily play an important role in tumorigenesis. HOXC9 and HOXD10 were reported playing critical roles in tumor progression in many malignant tumors. This study aimed to research the expression of HOXC9 and HOXD10 in papillary thyroid cancer, and to verify the prognostic and clinical significance of HOXC9 and HOXD10.MethodsImmunohistochemistry was used to determine the expression of HOXC9 and HOXD10 in 98 pairs of papillary thyroid cancer and paracancer tissues. Clinicopathologic data were collected and analyzed to verify the prognostic and clinical significance of HOXC9 and HOXD10.ResultsThe expression of HOXC9 and HOXD10 decreased in papillary thyroid cancer. The low expression of HOXC9 was associated with Hashimoto’s thyroiditis and lymph node metastasis (P<0.05). The low expression of HOXD10 was associated with extrathyroidal extension and lymph node metastasis (P<0.05). The co-expression rates of HOXC9 and HOXD10 was 44.90%. The low expression of both HOXC9 and HOXD10 was associated with lymph node metastasis (P<0.05).ConclusionsThe expression of HOXC9 and HOXD10 was downregulated in papillary thyroid cancer. Low expression of HOXC9 and HOXD10 might be related to the malignancy of papillary thyroid cancer. HOXC9 and HOXD10 may be used as diagnostic and prognostic biomarkers in the future.