Correlateions between UBE2T Expression and Immune Infiltration in

Different Cancers

Yunliang Yu

and Ting Li

Department of Medical Laboratory,

Yantai Affiliated Hospital of Binzhou Medical University,

Yantai, Shandong, 264100, China

Department of Pathology,

Yantai-Mountain Hospital of Yantai, Yantai Shandong, 264000, China

Keywords: UBE2T, Expression, Database, Prognosis, Lymphatic Metastasis, Immune Infiltration.

Abstract: Ubiquitin-conjugating Enzyme E2T (UBE2T) has been implicated in the development of several cancers.

However, the interaction of UBE2T and cancer immunity in many tumors remains uncertain. In this study,

we looked at the clinicopathological importance of UBE2T in various malignancies, as well as the links

between UBE2T and prognosis and tumor-infiltrating lymphocytes in a variety of cancers. The Gene

Expression Profiling Interactive Analysis (GEPIA) and Tumor Immune Estimation Resource (TIMER)

databases were used to extract UBE2T expression data. The Kaplan-Meier plotter and the GEPIA databases

were used to evaluate the relationship between UBE2T expression and cancer patients' clinical prognosis.

TIMER was used to look for relationships between UBE2T expression levels and tumor-infiltrating cells as

well as gene marker sets of immune infiltrates. The findings demonstrated that increased UBE2T expression

was substantially related to poor overall survival in individuals with breast cancer or lung adenocarcinoma.

In thymoma, stomach carcinoma, and ovarian cancer, patients with a higher UBE2T expression had a better

prognosis than those with low expression. Furthermore, elevated UBE2T expression impacted the prognosis

of lymphatic metastasis in individuals with breast, lung, and stomach cancer. There were clear positive or

negative relationships between UBE2T expression and immune cell infiltration in lung, stomach, and

thymoma cancers. Finally, UBE2T may be valued for its dual role in human malignancies because it may play

a significant role in the recruitment and modulation of immune infiltrating cells in tumors.

1 INTRODUCTION

Despite breakthroughs in early diagnosis and

intervention, cancer remains the leading cause of

death globally. It is generally recognized that tumor

immune microenvironment (TIME) influences cancer

patients' malignancy grade and prognosis (Julia

2017). Numerous studies have found that immune-

related systems play important roles in a variety of

human malignancies, and tumor immunotherapy has

changed cancer treatment (Gorabi 2020). Although

immunotherapeutic techniques are thought to be a

successful approach for cancer treatment,

immunotherapy using anti-PD-1 antibodies and anti-

PD-L1 demonstrated a partial response in advanced

lung and gastric cancer (Dermani 2019, Bernatchez

2013, Ninomiya 2018). Furthermore, the total

immunogenicity of gastric cancer is rather low, and

https://orcid.org/0000-0001-5450-160X

https://orcid.org/0000-0002-2140-2952

the efficacy of immunological therapy is rather

restricted (Lordick 2017). Because breast cancer is

also an immunostimulatory illness, some individuals

benefit from immunotherapy; nonetheless, it has

historically proved resistant to immunotherapy

(Pusztai 2016). As a result, it is crucial to investigate

the immunodepressive mechanism or

immunostimulatory functions in cancer patients in

order to uncover novel therapeutic targets for

immunotherapy of various tumors. Immune infiltrates

can influence tumor patients' prognosis and

responsiveness to treatment. Tumor infiltrating

lymphocytes (TILs) are commonly regarded as a

critical indicator of the immunological interaction

here between host and tumor, as well as possible

prognostic indicators of good or worse prognosis in

invasive malignancies (Baxevanis 2019).

Understanding the interplay between the tumor and

Yu, Y. and Li, T.

Correlateions between UBE2T Expression and Immune Inﬁltration in Different Cancers.

DOI: 10.5220/0011368400003444

In Proceedings of the 2nd Conference on Artiﬁcial Intelligence and Healthcare (CAIH 2021), pages 333-338

ISBN: 978-989-758-594-4

333

the host immune system is thus crucial for

discovering prognostic biomarkers, limiting

medication resistance, and creating novel diagnostic

methods.

Ubiquitin-conjugating Enzyme E2T (UBE2T)

was first identified in a case of Fanconi anemia.

UBE2T functions by combining with specifc E3

ubiquitin ligase to induce the degradation of

functional changes in substrate molecules. UBE2T is

increasingly recognized as a critical factor during

carcinogenesis in human nasopharyngeal, liver,

prostate, breast, gastric, lung cancers and so on.

Recent evidence also demonstrates that mutations or

impairment of the E2s can lead to severe disease

states, including chromosome instability syndromes,

cancer predisposition, and immunological disorders

(Ben-Neriah 2002). It has been observed that UBE2T

is increased in breast cancer and that suppressing

UBE2T expression can decrease breast cancer

cell invasion and metastasis. UBE2T was also shown

to be overexpressed in non-small cell lung cancer

(NSCLC) tissues and cell lines. In vitro, UBE2T

knockdown inhibited NSCLC cell proliferation,

migration, and invasion. It was also shown that

elevated UBE2T expression was associated with poor

differentiation and prognosis in patients with gastric

cancer. By modifying the expression of epithelial-

mesenchymal transition (EMT)-related factors,

suppressing UBE2T expression might reduce the

invasive and metastatic properties of gastric cancer

cells. Because a vast number of studies have shown

that inflammatory T cells, dendritic cells (DCs),

monocytes, and macrophages increase tumor

metastasis, it is possible that UBE2T is associated

with carcinogenesis and immune infiltration in many

malignancies.

The goal of this study is to look at the relationship

between UBE2T expression and the

clinicopathological importance of cancer patients in

databases including GEPIA, Kaplan-Meier plotter,

and TIMER. The relationship between UBE2T and

tumor-infiltrating immune cells in various tumor

microenvironments was also thoroughly investigated

utilizing the TIMER database. Furthermore, this

study looked at the link between the expression

pattern of UBE2T and various genes associated with

immune regulation in many malignancies for the first

time. This study not only demonstrated the

significance of UBE2T in malignancies, but it also

provided probable linkages and methods of

interaction between UBE2T and tumor immunity in

thymoma, breast, lung, and gastric cancers.

2 MATERIALS AND METHODS

2.1 GEPIA 2.0 Dataset

GEPIA (http://gepia.cancer-pku.cn/index.html) is an

interactive web site that analyzes RNA sequencing

expression data from 97,366, tumors and 8,587

normal samples from the TCGA (The Cancer

Genome Atlas) and the Genotype-Tissue Expression

(GTEx) projects. GEPIA was primarily employed in

this investigation to investigate the predictive

influence of UBE2T on overall survival (OS) and

disease-free survival (DFS) (DFS).

2.2 Kaplan Meier-Plotter Dataset

The Kaplan Meier-plotter database can examine

predictive values of gene mRNA expression in

patients with breast, gastric, lung, and ovarian cancer,

as well as miRNA expression in patients with liver

and breast cancer (http://kmplot.com/analysis/).

Using the Kaplan Meier-plotter database, we

investigated the relationship between UBE2T

expression and prognosis in breast, lung, stomach,

and ovarian cancer. By comparing the E-MTAB-365

and Gene Expression Omnibus (GEO) datasets, we

also confirmed whether UBE2T expression impacts

the prognosis of breast, lung, and gastric cancer in

patients with lymphatic metastases (dataset:

GSE30219 and GSE62254). The Kaplan Meier-

plotter dataset included the UBE2T expression and

OS rates, RFS (relapse-free survival) rates,

progression-free survival (PFS) rates and post-

progression survival (PPS) rates of 426 patients with

breast cancer (dataset: E-MTAB-365), 307 patients

with lung cancer (dataset: GSE30219), 300 patients

with gastric cancer (dataset: GSE62254) and 285

patients with ovarian cancer (dataset: GSE9891).

2.3 TIMER Database Analysis

TIMER is an online resource server for systematic

analyses of tumor-infiltrating immune cells in various

cancer types (https://cistrome.shinyapps.io/timer/).

The TIMER platform was utilized in this study to

detect distinct profiles of immune cells and to

calculate the immunological estimate of the TCGA

dataset. These immune cells include B cells, CD4+ T

cells, CD8+ T cells, neutrophils, macrophages, and

DCs. The gene module on TIMER demonstrated a

link between UBE2T expression and varied degrees

of immune infiltration. Furthermore, correlations

between UBE2T expression and gene markers of

tumor-infiltrating immune cells were also explored

CAIH 2021 - Conference on Artiﬁcial Intelligence and Healthcare

334

by correlation modules. Correlation modules were

also used to investigate connections between UBE2T

expression and gene biomarkers of tumor-infiltrating

immune cells. The correlation method generates

expression scatter plots between two user-defined

genes in a specific cancer type, where the x-axis

indicates relevant marker genes and the left y-axis

represents UBE2T. For all correlation graphical

representations, Spearman rank correlation was

employed.

2.4 Statistical Analysis

A one-sided Student's t-test was used to obtain P-

values and fold-changes for UBE2T expression from

the GEPIA database. GEPIA and Kaplan-Meier

survival plots were used to assess the survival rates

estimated from the date of diagnosis. The outcomes

of Kaplan-Meier plots and GEPIA are represented as

Hazard ratios (HR) and P-values or Cox P-values

from a log-rank test. Because associations can be

positive or negative, the absolute values of the

correlation coefficients in TIMER were used to

determine the strength of Spearman rank correlation.

A statistically significant difference was defined as

one with a value of P<0.05.

3 RESULTS

3.1 UBE2T Gene Was Ubiquitously

Overexpressed in Tumor Tissue

TIMER Database Analysis

Using TIMER-generated RNA-sequencing (RNA-

Seq) data from multiple malignant tumors in the

TCGA, we investigated at UBE2T expression. The

UBE2T mRNA expression was proven to increase

dramatically in BLCA (bladder urothelial

carcinoma), BRCA (breast invasive cancer), CHOL

(cholangio carcinoma), COAD (colon

adenocarcinoma), ESCA (esophageal carcinoma),

HNSC (head and neck cancer), KICH (kidney

chromophobe), KIRC (kidney renal clear cell

carcinoma), KIRP (kidney renal papillary cell

carcinoma), LIHC (liver hepatocellular carcinoma),

LUAD (lung adenocarcinoma), LUSC (lung

squamous cell carcinoma), PRAD (prostate

adenocarcinoma), READ (rectum adenocarcinoma),

STAD (stomach adenocarcinoma), THCA (thyroid

carcinoma) and UCEC (uterine corpus endometrial

carcinoma) compared with adjacent normal tissues.

We also discovered that UBE2T was widely

expressed in practically all cancer types, including

breast, lung, ovarian, and stomach cancers, after

digging further into the GEPIA database. In the same

way, UBE2T expression was elevated in distinct

BRCA subtypes, LUAD, and LUSC.

3.2 High UBE2T Expression Was

Significantly Related to the

Prognosis of Tumors

The GEPIA database was used to investigate the links

between UBE2T expression and cancer patient

prognosis. In many common cancer types, higher

expression of UBE2T was associated with a poorer or

better survival rate, including ACC (adrenocortical

carcinoma), DLBC (diffuse large B-cell lymphoma),

KICH (kidney chromophobe), KIRC, KIRP, LGG

(lower grade glioma), LIHC, MESO (mesothelioma),

PAAD (pancreatic adenocarcino (uveal melanoma).

Among them, we found that high UBE2T expression

was marginally associated with poor prognosis in

BRCA (OS Logrank P=0.017) and LUAD (OS

Logrank P=0.001; DFS Logrank P=0.0091). There

was no significant relevance between UBE2T

expression and prognosis of cancer patients in LUSC.

However, high expression level of UBE2T was

associated with better prognosis in STAD (DFS

Logrank P = 0.04), THYM (thymoma) (OS Logrank

P=0.005) and OV (ovarian serous

cystadenocarcinoma) (OS Logrank P=0.011). Such

differences are suggestive of UBE2T may

demonstrate a dual role in the malignant tumor

development process. Meanwhile, high expression of

UBE2T was found to be related to different tumor

stages in BRCA (P=3.34e-5), LUAD (P=0.00164)

and OV (P=0.0107). Nevertheless, this correlation

was not found in LUSC or STAD.

3.3 High Expression of UBE2T

Influences the Prognosis of Breast,

Lung and Gastric Cancer in

Patients with Lymphatic Metastasis

We used the Kaplan Meier-plotter database to study

the predictive functions of UBE2T in breast, lung,

and gastric cancer patients in attempt to comprehend

the independent influence of UBE2T expression on

survival. We discovered that greater UBE2T mRNA

expression was linked to a poorer RFS in breast

cancer (P=0.0028) and lung cancer (Logrank P=3.3e-

7). However, the significant of UBE2T mRNA levels

were associated with better OS and PPS in gastric

cancer (OS Logrank P=3.7e-5; PPS Logrank P=1.4e-

Correlateions between UBE2T Expression and Immune Inﬁltration in Different Cancers

335

6) and ovarian cancer (PFS Logrank P=0.011). These

results are consistent with our above results.

In the TNM staging system, the N category relates

to lymph node involvement; N0 indicates no regional

lymph node metastasis, whereas N1-N3 indicate

regional lymph node metastasis. We discovered that

greater UBE2T expression was linked with poorer

RFS in lymph node positive breast cancer patients

(P=0.0043), but not with RFS in lymph node negative

cancer patients. In lung cancer patients, increased

UBE2T mRNA expression was linked with poorer

OS in stage N0 (P=0.0067) and stage N2 (P=0.033),

but not with OS in stage N1 (P=0.2343). In gastric

cancer, elevated UBE2T mRNA expression was

linked with a better prognosis in stage N0 (OS

P=0.0017; PPS P=0.001) and stage N1 (OS P=0.002;

PPS P=0.0011) patients but not in stage N2 (OS

P=0.153; PPS P=0.13) and stage N3 (OS P=0.2084;

PPS P=0.079) patients. To summarize, our findings

suggest that elevated UBE2T expression may affect

the prognosis of patients with lymph node metastases

in breast, lung, and gastric cancer.

3.4 UBE2T Expression Is Correlated

with Immune Infiltration Level in

LUAD, STAD And THYM

We predicted that because tumor-infiltrating

lymphocytes are an independent predictor of lymph

node status and prognosis in malignancies, immunity

in tumors may be connected to UBE2T expression.

We looked at the relationships between UBE2T

expression and TIL quantity as measured by tumor

purity. TIMER examined the relationship between

UBE2T expression and immune infiltration levels in

39 cancer types. Higher UBEB2T expression was

shown to be substantially linked with tumor purity in

27 different forms of cancer. Furthermore, patients

with greater UBE2T expression demonstrated

considerable infiltration of CD8+ cells in 7 cancer

types, CD4+ T cells in 28 cancer types, macrophages

in 25 cancer types, neutrophils in 18 cancer types, and

DCs in 19 cancer types. Among these, UBE2T

expression is linked to varying amounts of immune

infiltration and tumor purity in BRCA, LUAD,

STAD, and OV. Furthermore, in breast cancer, there

were very moderate relationships between UBE2T

expression and infiltrating quantities of B cells and

CD4+ T cells. The degree of UBE2T expression

infiltrating B cells, CD4+ T cells, macrophages,

neutrophils, and DCs revealed significant negative

correlations in LUAD. B cells, CD8+ cells, CD4+ T

cells, macrophages, neutrophils, and DCs infiltrated

less frequently in UBE2T overexpression instances in

STAD. However, in ovarian cancer, there was almost

no association between UBE2T expression and

immune cell infiltration levels. Surprisingly, the

infiltrating levels of B cells, CD8+ T cells, CD4+ T

cells, macrophages, and DCs have higher substantial

positive relationships with UBE2T expression levels

in THYM.

In general, this then implies that UBE2T may be

implicated in immune infiltration in thymoma, lung

and gastric cancer, especially those of CD4+T cells,

macrophages and DCs. UBE2T may play important

roles in the cross-talk between tumor and immune

environment in order to drive tumorigenesis.

3.5 UBE2T Is Correlated with Typical

Immune Marker Genes in LUAD

and STAD

We used the TIMER database to show the

relationships between UBE2T and the immunological

marker sets of diverse immune cells from LUAD and

STAD. These immune cells include CD8+ T cells, T

cells (general), B cells, monocytes, tumor associated

macrophages (TAMs), neutrophils, natural killer

(NK) cells, and DCs, which contribute to the

immunosuppressive or immunomodulatory features

of the tumor microenvironment. CD4 T cells may

develop into a variety of helper and regulatory T-cell

lineages, including Th1, Th2, Th9, Th17, TFH, and

CD4+Foxp3+ Tregs. In this work, we looked at

several types of functional T cells, such as Th1 cells,

Th2 cells, and Tregs. We discovered that STAT6

expression levels in T cells had a somewhat

unfavorable connection with UBE2T expression in

LUAD. Furthermore, a somewhat favorable

connection between UBE2T and GZMB of T cell

fatigue was discovered.

3.6 Correlations between UBE2T and

Immune Marker Genes in THYM

In THYM, the correlations between UBE2T and

immune marker sets of T cells are complex. There

existed strong positive correlations between UBE2T

and the expression levels of CD8A (P=1.37e-18,

r=0.815) and CD8B (P=5.49e-26, r=0.782) of CD8+T

cells, CD2 (P=4.04e-26, r=0.793), CD3D (P=8.44e-

20, r=0.722) and CD3E (P=1.06e-30, r=0.832) of T

cell (general), GATA3 (P=3.15e-21, r=0.74) of Th2

cells, PD-L1 (PDCD1) (P=1.79e-15, r=0.656) of T

cell exhaustion in THYM. However, significant

negative correlations were identified between

UBE2T and the expression levels of STAT4

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336

(P=2.26e-4, r=-0.337), TNF (P=4.38e-9, r=-0.514)

and TNF-α (IFNG) (P=5.28e-7, r=-0.448) of Th1

cells, STAT5A (P=9.16e-5, r=-0.357) and STAT6

(P=7.63e-4, r=-0.31) of Th2 cells, STAT3 (P=1.11e-

5, r=-0.397) of Th17 cells, FOXP3 (P=6.17e-6, r=-

0.408) of Tregs, CTLA4 (P=5.51e-6, r=-0.409) of T

cell exhaustion in THYM.

4 CONCLUSIONS

Lung cancer is widely recognized as one of the most

common malignant tumors and the main cause of

cancer-related mortality worldwide. Similarly, breast

cancer still has the highest fatality rate among

gynecological cancers due to widespread

intraperitoneal metastases at an early stage and a lack

of effective early detection measures. Despite the fact

that immunotherapy has been shown to be effective

in the treatment of advanced malignancies, cancer

immunotherapy has a number of adverse effects as

well as other drawbacks. Therefore, there is an urgent

requirement for the identification of new targets for

immunotherapy for these cancers.

One of the most ubiquitous regulatory processes

in all eukaryotes is protein modification via covalent

attachment of ubiquitin (Dermani 2019). Ubiquitin

and ubiquitin-like proteins modify proteins, which is

a fundamental regulatory step in the innate and

adaptive immune responses. UBE2T was discovered

to be a critical regulator and oncogene in numerous

malignancies, including breast, lung, prostate, and

bladder cancer, as a member of the ubiquitin-

conjugating E2 family in the ubiquitin-proteasome

system.Many immune signaling pathways depend on

ubiquitin chain production, and the ubiquitin

conjugating enzyme is important in immune receptor

signaling (Dermani 2019). On the other hand, the

relationship between UBE2T expression and immune

infiltration levels, on the other hand, has never been

detected before. This study initially looked at the

levels of UBE2T expression in distinct cancer types

using publicly available information. The majority of

cancer types, including thymoma, breast, lung,

gastric, and ovarian cancer, have elevated UBE2T

expression. Following that, we investigated the

correlations between UBE2T expression and clinical

indications. We discovered that elevated UBE2T

expression was associated with a poor outcome in

breast and lung cancer. The outcomes for gastric

cancer, thymoma, and ovarian cancer, on the other

hand, were diametrically opposed. Patients with

greater levels of UBE2T expression had a better

prognosis. Furthermore, the expression of UBE2T

has been linked to the prognosis of individuals with

lymph node metastases in breast, lung, and gastric

cancer. Further research revealed that high UBE2T

levels were, to some extent, associated with tumor

purity in LUAD and STAD. The interplay of tumor

stroma and tumor-infiltrating lymphocytes has a

significant impact on cancer patient outcomes. In

LUAD and STAD, we also discovered negative

relationships between the degree of UBE2T

expression and the infiltration of CD4+ T cells, DCs,

and macrophages. THYM, on the other hand,

revealed strikingly positive relationships. This

implies that UBE2T in LUAD may have

immunoregulatory and cancer-promoting

characteristics. While STAD and THYM may have

sophisticated mechanisms for anticancer activity, It

should be mentioned that the link between UBE2T

expression and the prognosis of gastric cancer

patients contradicted prior research findings. The

inconsistent results may be due to differences in the

techniques used or differences in the tumor samples

examined. Thus, further study is needed to validate

our findings and hypothesis.

Based on the findings, we hypothesized that

UBE2T may have a role in tumor immunology

regulation. So we went a step further and used the

TIMER database to find a link between UBE2T

expression and immune cell marker genes in LUAD,

STAD, and THYM. T cells are exceedingly

complicated and diverse in vivo, with continual

renewal. We initially looked for links between

UBE2T and T-cell marker genes. THYM discovered

strong positive connections between UBE2T and

CD8A and CD8B of CD8+ T cells, as well as CD2,

CD3D, and CD3E of general T cells. This suggests

that increased UBE2T expression may be linked to

the recruitment of CD8+ T cells and general T cells

in THYM. GATA3, a zinc-finger transcription factor

in the GATA family, is recognized to be a critical

regulator of Th2 development. In THYM, we

discovered a substantial positive connection between

UBE2T expression and GATA3. These findings

suggest that UBE2T may influence Th2 cell

development. STAT6 is a downstream effector of

Th2 cytokine signaling, and high STAT6 levels

indicate a robust T-helper 2 type immunological

response (Julia 2017). This study found a substantial

negative connection between UBE2T expression and

STAT6, indicating that UBE2T is involved in the T-

helper type 2 response in LUAD and TYHM. In

THYM, there was a strong positive connection

between UBE2T expression and PD-L1 (Julia 2017).

Furthermore, in LUAD, GZMB is modestly positive

correlated with UBE2T expression, but CTLA4 is

Correlateions between UBE2T Expression and Immune Inﬁltration in Different Cancers

337

strongly negatively correlated with UBE2T

expression in THYM. T-cell exhaustion is recognized

to contribute to disease development since the

immune system's ability to control an infection or

tumor decreases as T-cell functioning declines. Our

findings suggest that UBE2T may play a role in T cell

fatigue in a variety of malignancies. In summary,

these findings indicate that UBE2T is involved in T-

cell function control and that UBE2T may play a

variety of roles in various cancers.

To conclude, UBE2T may play a dual function in

several human malignancies, including promoting

and preventing carcinogenesis. UBE2T expression is

linked to prognosis and immune infiltration levels in

CD4+ T cells, CD8+ T cells, neutrophils,

macrophages, and DCs from many malignancies,

including LUAD, STAD, and THYM. The effects of

the UBE2T bidirectional interaction in promoting or

inhibiting a tumor phenotype are unknown but should

be investigated.

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