LINC00641/miR-378a and Their Cross-Talk with TNF-α/IFN-γ as Potential Biomarkers in Ulcerative Colitis and Crohn's Diseases

Abstract

The most well-known forms of inflammatory bowel disease (IBD) that affect the entire gastrointestinal tract are ulcerative colitis (UC) and Crohn's disease (CD). The serum profile of inflammatory biomarkers and noncoding RNA and their role in the propagation of the inflammatory process remains controversial. Thus, this study was designed to examine the relationship between hematological profile, C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), interferon-gamma (INF-γ), and the expression of LINC00641 and miR-378a in individuals with IBDs. In addition, we elucidated the correlation between the expression of LINC00641 and miR-378a and the biochemical variables analyzed. This retrospective study analyzed 94 unrelated participants. Group I included healthy controls, Group II consisted of participants diagnosed with UC, and Group III consisted of participants diagnosed with CD. Patients with IBDs experienced significant elevations in CRP, total leukocyte count, platelets, erythrocyte sedimentation rate, TNF-α, and INF-γ. However, participants with IBD had lower hemoglobin and albumin levels than healthy control participants. Moreover, the expression levels of LINC00641 and miR-378a were elevated in participants with IBD, with a significant difference between participants with IBD and healthy controls. The most striking observation was a clear association between serum LINC00641 and miR-378a levels and the biochemical variables assessed. This study demonstrated a positive correlation between the expression of LINC00641/miR-378a and TNF-α in patients with UC and CD patients. This study suggests that LINC00641 and miR-378a are prospective biomarkers and noninvasive screening tools for IBDs, which may help predict the progression of complications.

Introduction

Inflammatory bowel disease (IBD), a lifelong disease, has been a global health issue with a steadily increasing incidence in almost 7 million people worldwide (Kaplan and Windsor, 2021). The most well-known forms of IBD are ulcerative colitis (UC) and Crohn's disease (CD), which affect the entire gastrointestinal tract and frequently cause irreversible gut damage requiring hospitalization or medical procedures (Kaplan and Windsor, 2021; Lamb et al., 2019).

UC is an idiopathic and emerging risk factor for inflammation that affects the colon. The hallmark of UC is chronic progressive intestinal inflammation (Khalil et al., 2023; Lamb et al., 2019). CD frequently occurs anywhere in the digestive tract from the mouth to the anus (Baumgart and Carding, 2007; Baumgart and Sandborn, 2012). Prompt diagnosis of IBDs is crucial for its management and alleviation by improving prognosis and therapy (Matsuoka et al., 2018). This opens new avenues for exploring and scrutinizing novel candidate biomarkers for IBD diagnosis.

Tumor necrosis factor-alpha (TNF-α) and interferon-gamma (INF-γ), two essential proinflammatory cytokines in the pathogenesis of IBD, are associated with disease characteristics (Hiroshi et al., 2022). Proinflammatory cytokines are produced in excessive amounts in inflammatory situations, which not only enhance the host organism's immune response but also have negative impacts that can affect metabolic control and hemodynamic regulation (Jarczak and Nierhaus, 2022). Thus, inhibiting TNF-α and INF-γ is effective in causing and maintaining clinical reactions and IBD remission.

Long noncoding RNAs (lncRNAs) are noncoding RNA that encompass ∼200–600 nucleotides. However, they also regulate gene expression through many meaningful regulatory procedures (Ayeldeen et al., 2023; Statello et al., 2021). LINC00641 acts as an anticancer RNA, inhibiting bladder cancer progression by regulating miR-197-3p14 (Han and Zhang, 2022).

MicroRNAs (miRNAs) represent a noncoding form of RNA associated with biologic process varieties and modulate the expression of target genes (O'Brien et al., 2018). Furthermore, miRNAs are linked to several metabolic pathways and are used to diagnose diseases such as colorectal cancer and breast cancer (Chen et al., 2019; Khaliefa et al., 2023; Si et al., 2019). miR-378a contributes to various metabolic pathways by inhibiting the Wnt/β-catenin signaling pathway, which in turn inhibits cancer cells (Deldar et al., 2021; Yu et al., 2021).

To our knowledge, the differential expression of LINC00641 and miR-378a in individuals with IBDs and their correlation with C-reactive protein (CRP), TNF-α/INF-γ, and hematological profiles have not been explored. Thus, this study was designed to scrutinize the relationship between hematological profile, CRP, TNF-α/INF-γ, and the expression of LINC00641 and miR-378a in individuals with IBDs and explore their correlations and diagnostic potential for IBDs.

Materials and Methods

In this retrospective case–control study, 94 unrelated participants were evaluated during the period from March 2022 to January 2023, and written informed consent was provided for participation before enrollment in the study. Group I included healthy controls, Group II consisted of participants diagnosed with UC, and Group III consisted of participants diagnosed with CD. Endoscopy and clinical criteria were used in the diagnosis and differentiation of UC and CD. This retrospective study followed the ethical principles of the Declaration of Helsinki and the agreement of the Research and Ethics Committee (FMBSUREC/02102022/Abd-elhameed). Patients with acute or chronic renal failure, cardiac disease, cirrhosis, malignancy, or acute or chronic infections were excluded from the study.

All participants had fasting venous blood samples (10 mL) taken for routine analysis, including the hematology profile, CRP, and liver profile, and were stored at −80°C until LINC00641 and miR-378a expression analysis.

TNF-α and INF-γ assay

Serum TNF-α measurement was carried out using TNF-α ELISA kits provided by Avi-Bion (Orgenium Laboratories, Finland), according to Corti et al. (1992). Serum INF-γ were determined by using a commercially available enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, Inc., Minneapolis, MN) according to the manufacturer's instructions.

LINC00641 and miR-378a expression analysis

TRIzol reagent (Invitrogen) was used to isolate total RNA encoding miRNAs and lncRNAs from serum samples. A NanoDrop^® (ND)-1000 spectrophotometer (NanoDrop Technologies, Inc.) was used to assess and evaluate isolated RNA. Using the miScript II RT kit (Qiagen, Valencia, CA), whole RNA was reverse transcribed in a final volume of 20 μL RT reactions. Primers specific for LINC00641 and miR-378a were used to amplify the cDNA using miScript SYBR Green Master Mix.

The amplification data were assessed using SNORD 68 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the housekeeping genes. Differential expression was assessed using the 2^−ΔΔCt method (Livak and Schmittgen, 2001).

Statistical analysis

SPSS statistical software, version 22, was used to evaluate the data. Statistical data were expressed as mean ± standard error of the mean and percentages. One-way analysis of variance (ANOVA) was used to investigate differences between the groups under consideration. Post hoc multiple comparisons tests (Duncan) at P < 0.05 were used for pairwise comparisons. In addition, the correlations among the analyzed variables were assessed using Pearson's correlation coefficient, and diagnostic performance tests were determined using receiver operating characteristic (ROC) curve analysis.

Results

In this study, we included 30 participants with UC, 34 with CD, and 30 healthy individuals; the demographic and clinical data of the study participants are given in Table 1.

Table 1.

Demographic Data and Clinical-Pathological Features of the Studied Groups

Variables	Healthy control (n = 30)	UC subjects (n = 30)	CD subjects (n = 34)
Age (mean ± SD) (range)	29.13 ± 6.97 (18–42)	31.93 ± 10.68 (18–55)	33.12 ± 11.94 (17–62)
Gender
Male, n (%)	14	19	21
Female, n (%)	16	11	13
Extent
Left side		13
Pancolitis		14
Procolitis		3
L1 (ileum)			10
L2 (Colon)			10
L3 (Colon+ileum)			14
CDAI (mean ± SE)		———	271.62 ± 22.52

CD, Crohn's disease; CDAI, Crohn's disease activity index; UC, ulcerative colitis.

Regarding the hematological and biochemical profiles, all participants with UC and CD had significantly higher rates of total leukocyte count (TLC), PLTs, and erythrocyte sedimentation rate (ESR) than healthy individuals (P < 0.05). However, the hemoglobin (Hb) and albumin levels in participants with UC and CD were considerably lower than those in healthy subjects. Participants with UC and CD showed elevated CRP levels compared with healthy participants (Table 2).

Table 2.

Hematological and Biochemical Characteristics of Studied Participants

Variables	Healthy control	UC	CD
Hb	12.59 ± 0.23^b	11.88 ± 0.38^ab	11.23 ± 0.26^a
TLC	4.59 ± 0.13^a	7.49 ± 0.34^b	8.45 ± 0.54^b
Platelets	299.27 ± 1.75^a	324.43 ± 1.11^a	376.85 ± 1.77^b
ESR	5.94 ± 0.35^a	31.53 ± 0.58^b	35.44 ± 0.88^b
CRP	2.57 ± 0.18^a	9.23 ± 0.43^b	16.60 ± 0.26^c
Albumin	4.61 ± 0.04^b	3.75 ± 0.15^a	3.53 ± 0.13^a

Data are expressed as mean ± SEM; According to the Duncan multiple range test, different letters indicate statistical significance different means.

CRP, C-reactive protein; ESR, erythrocyte sedimentation rate; Hb, hemoglobin; SEM, standard error of the mean; TLC; total leukocyte count.

To determine whether LINC00641 and miR-378a were differentially expressed in the serum of UC, CD, and control individuals, their expression was measured using quantitative real-time polymerase chain reaction. In individuals with UC, the expression level of miR-378a and LINC00641 were elevated compared with that of healthy controls. Similarly, CD patients exhibited even higher levels of expression of these markers than UC patients. Furthermore, it is worth mentioning that serum TNF-α and INF-γ expression levels were significantly (P < 0.001) elevated in both UC and CD patients compared with healthy participants. TNF-α and INF-γ expression levels demonstrated a noticeable elevation in patients with CD versus those with UC (Table 3).

Table 3.

Expression Levels of LNIC00641, miR-378a, TNF-α, and INF-γ Regarding UC and CD Patients

Variables	Control	UC	CD
LINC00641	0.98 ± 0.01^a	1.49 ± 0.16^a	4.60 ± 0.49^b
miR-378a	1.03 ± 0.01^a	5.60 ± 0.30^b	12.46 ± 0.72^c
TNF-α (pg/mL)	15.87 ± 0.41^a	30.09 ± 1.00^b	34.15 ± 1.72^c
INF-γ (pg/mL)	16.70 ± 0.59^a	46.82 ± 2.34^b	82.17 ± 4.20^c

Data are expressed as mean ± SE. According to the Duncan multiple range test, different letters indicate statistical significance different means.

TNF-α, tumor necrosis factor-alpha.

There were no significant differences (P > 0.05) in the expression levels of LINC00641 and miR-378a among UC patients with pancolitis, left-sided, and proctitis involvement (Table 4). In patients with CD, there were significant differences in the expression levels of LINCOO641 and miR-378a in patients with CD depending on the extent of the disease by colonoscopy (Table 5).

Table 4.

Differential Expression Levels of LNIC00641, and miR-378a Among UC Patient in Relation to the Extent of Disease by Colonoscopy

Extent of disease by colonoscopy	LINC00641	miR-378a
Pancolitis	1.412 ± 0.179^a	5.399 ± 0.451^a
Left-sided	1.445 ± 0.274^a	5.688 ± 0.501^a
Procolitis	2.0553 ± 1.00^a	6.227 ± 0.650^a
F value	0.639	0.324
P value	>0.05	>0.05

Data are expressed as mean ± SE. According to the Duncan multiple range test, different letters indicate statistical significance different means.

Table 5.

Differential Expression Levels of LNIC00641, and miR-378a Among CD Patients in Relation to the Extent of Disease by Colonoscopy

Extent of disease by colonoscopy	LINC00641	miR-378a
Ileum	5.690 ± 0.972^b	11.273 ± 1.710^a
Colon	5.528 ± 0.926^b	15.237 ± 0.724^b
Colon+ileum	3.164 ± 0.550^a	11.320 ± 0.943^a
F value	3.520	3.584
P value	<0.05	<0.05

Data are expressed as mean ± SE. According to the Duncan multiple range test, different letters indicate statistical significance different means.

Pearson correlation analysis was performed to determine the relationship between LINC00641/miR-378 expression and the biochemical parameters of UC patients (Table 6). According to correlation analysis, miR-378a had substantial positive relationships with LINC00641, TNF-α, INF-γ, CRP, and TLC levels. Moreover, no statistical significance was observed between LINC00641 and TNF-α, INF-γ, CRP, or TLC levels (P > 0.05). In addition, miR-378/LINC00641 had a nonsignificant negative relationship with Hb levels in patients with UC (Table 6).

Table 6.

Pearson Correlation Coefficient Between Biochemical Variables in Study Participants

Parameter		Biochemical variables	UC (n = 30)	CD (n = 34)
FC miR-378		LINC00641
	r		0.339^**	0.640^**
	P		<0.001	<0.001
		TNF-α
	r		0.716^**	0.772^**
	P		<0.001	<0.001
		INF-γ
	r		0.763^**	0.637^**
	P		<0.001	<0.001
		CRP
	r		0.587^**	0.579^**
	P		<0.001	<0.001
		Hb
	r		−0.219	−0.348^**
	P		0.09	0.005
		TLC
	r		0.681^**	0.551^**
	P		<0.001	<0.001

FC LINC00641		miR-378
	r		0.339^**	0.640^**
	P		<0.001	<0.001
		TNF-α
	r		0.154	0.600^**
	P		>0.05	<0.001
		INF-γ
	r		0.202	0.530^**
	P		>0.05	<0.001
		CRP
	r		0.171	0.406^**
	P		>0.05	0.001
		Hb
	r		−0.022	−0.288^*
	P		>0.05	0.021
		TLC
	r		0.161	0.285^*
	P		>0.05	0.022

Significant linear correlation at P < 0.01 (2-tailed).

Significant linear correlation at P < 0.05 (2-tailed).

There was correlation between LINC00641/miR-378 expression and biochemical parameters of CD patients. According to correlation analysis, LINC00641 and miR-378 expression had substantial positive relationships with TNF-α, INF-γ, CRP, and TLC levels. In addition, miR-378/LINC00641 had a significant negative relationship with Hb levels in patients with UC (Table 6).

Diagnostic performance of serum LINC00641, and miRNA-378a

With an area under the ROC curve (AUC) of 0.843, serum LINC00641 was able to distinguish healthy controls from UC in ROC curve analysis, with confidence interval (CI) = 0.737–0.949, P < 0.001, sensitivity = 63.33%, specificity = 100% (Fig. 1a). Serum LINC00641 discriminated healthy controls from CD patients with AUC = 0.92, 95% CI = 0.836–1.004, with sensitivity of 88.24%, and 100% specificity (Fig. 1b). Also, serum LINC00641 discriminated CD from UC with AUC = 0.839, 95% CI = 0.734–0.943, P < 0.001, with sensitivity = 79.41%, specificity = 83.33% (Fig. 1c).

FIG. 1.

ROC analysis regarding LINC00641 between (a) UC and healthy control participants. (b) CD and healthy control participants. (c) Crohn's disease and UC patients. CD, Crohn's disease; ROC, receiver operating characteristic; UC, ulcerative colitis.

On the contrary, with an AUC of 0.968, miR-378a discriminated healthy controls from UC with 95% CI = 0.906–1.030, P < 0.001, 96.67% sensitivity, and 100% specificity (Fig. 2a). Moreover, with an AUC of 0.974, serum miR-378a distinguished controls from patients with CD with 95% CI = 0.914–1.027, P < 0.001, 97.06% sensitivity, and 100% specificity (Fig. 2a). In addition, it distinguished from UC with AUC = 0.922, 95% CI = 0.836–1.007, P < 0.001, with sensitivity = 91.18% and specificity = 100% (Fig. 2c).

FIG. 2.

ROC analysis regarding miR-378a between (a) UC and healthy control participants. (b) CD and healthy control participants. (c) CD and UC patients.

Discussion

Patients with IBD are predisposed to an assortment of problems owing to inadequate management and control of inflammation. This opens new avenues for exploring and scrutinizing novel candidate biomarkers for early IBD diagnosis. Accordingly, this study sought to demonstrate the differential expression of LINC00641 and miR-378 and their correlation in patients with IBD (UC and CD).

Although the exact pathogenicity of IBD forms remains unknown, it is well established that both arise as a consequence of genetic predisposition and immune gut flora dysregulation (Dalal and Chang, 2014). However, localization and endoscopic observations are not shared between both forms of IBD (Vespa et al., 2022).

In this study, white blood cell, platelet, ESR, and CRP levels were noticeably elevated in participants with UC and CD compared with healthy controls. Our results are consistent with those of Sayar et al. (2020) and Chen et al. (2020), who reported that patients with IBD exhibited a noticeable elevation in hematological profiles and CRP levels.

The blood expression of CRP is the most comprehensive biomarker of inflammatory disorders. The recurrence of IBD seems to be attributed to clinical signs and abnormally high CRP expression (Sproston and Ashworth, 2018). Furthermore, Karoui et al. (2011) explored the link between clinical and endoscopic activity and CRP level in patients with UC. However, high CRP levels are not disease specific, and they can occur in a variety of circumstances, including non-IBD enteritis, tissue damage, diabetes, malignancies, and cardiovascular disease (Fu et al., 2020; Hart et al., 2020; Jeong et al., 2019).

Our study revealed a significant decline in Hb and albumin levels in patients with UC and CD. Accordingly, previous studies have reported a noticeable decline in Hb and albumin levels in patients with UC and CD (Khan et al., 2017). In addition, Woźniak et al. (2019) stated that hypoalbuminemia has a significant impact on the onset of anemia in patients with IBD, and the severity of the inflammatory response is inversely correlated to albumin levels that arise from a decrease in albumin synthesis in the liver (Dalal and Chang, 2014). Moreover, Levitt and Levitt (2016) reported that hypoalbuminemia was correlated with disease activity, nutritional status, unresponsiveness to treatment, and an increased risk of colectomy in patients with acute severe UC.

This study demonstrated a statistically significant elevation in TNF-α and INF-γ levels in UC and CD patients compared with healthy participants. These data are consistent with their enhanced expression previously reported in patients with IBDs, which supports their role in the pathogenesis of IBDs (Eltayeb et al., 2020). Our findings are also supported by other studies investigating TNF-α levels in patients with IBD (Eltayeb et al., 2017). Increased TNF-α and INF-γ levels in the blood of UC and CD patients, released mainly from the intestinal colonic epithelium, have been proposed as markers of inflammation that contribute to disease severity (Eltayeb et al., 2020).

In this study, all participants with UC and CD showed elevated LINC00641 expression levels. In addition, patients with CD exhibited noticeably higher LINC00641 expression than those with UC. In concordance, Padua et al. (2016) and Chen et al. (2016), revealed that the expression of LncRNAs exhibited a noticeable overexpression in IBDs and was linked with the severity, intestinal permeability, and cell-intrinsic functions such as apoptosis and proliferation. Similarly, Hu et al. (2020) reported that LINC00641 is a new potential biomarker in patients with gastric adenocarcinoma, providing a predictive regimen for global survival.

Concerning the expression of miR-378a, all patients with UC and CD exhibited a notable elevation of miR-378a expression. Moreover, the differential expression of miR-378a was significantly higher in patients with CD than in those with UC. Consistent with our results, previous investigations have demonstrated an alteration in miR-378a expression in patients (Dubois-Camacho et al., 2019; Duttagupta et al., 2012). These alterations may be owing to the small number of patients included in previous studies or the use of pharmacological agents by the participants, which could reflect treatment rather than disease effects.

In our study, according to Pearson's correlations, serum LINC00641, miR-378a, CRP, TNF-α, INF-γ, and TLCs were positively associated. Moreover, the serum expression of LINC00641 and miR-378a was negatively associated with Hb levels in the study participants. Consistent with our results, Alper et al. (2017) and Solem et al. (2005) showed a positive correlation between CRP elevation and the TLC. Wang et al. (2016) reported a positive correlation between serum levels of miR-223, ESR, and CRP as indicators of disease activity in UC.

An ROC curve was constructed to test the diagnostic ability of LINC00641, producing a diagnostic accuracy (AUC = 92%) and a sensitivity = 88.24% and 100% specificity in discriminating patients with CD from healthy control participants. miR-378a had a high diagnostic accuracy (AUC = 97.1%) and a 97.06% sensitivity, specificity of 100% for distinguishing healthy controls from CD patients. It had a respectable capability to discriminate between CD and UC (AUC = 92.2%) and 91.18% sensitivity.

Conclusion

To our knowledge, this is the first study to report the differential expression of LINC00641 and miR-378a in individuals with IBDs and their correlation with CRP, hematological profile (ie, Hb and TLC), TNF-α, and INF-γ. This study showed that the elevation of LINC00641 and miR-378a coincided with the elevation of TNF-α and INF-γ in patients with IBD. This suggests that LINC00641 and miR-378a are prospective biomarkers of IBDs, which may help predict the progression of complications. However, this study has some limitations, including the sample size power analysis and the differential expression of miR-378a and LINC00641 in treatment response variability to TNF-α inhibitors. Larger and well-designed studies are required to further confirm the exact roles of LINC00641 and miR-378a in IBD development, progression, and treatment response to TNF-α inhibitors.

Footnotes

Authors' Contributions

N.A.H. and O.G.S. conceived and designed the research study; N.A.A-.H., O.G.S. contributed reagents/materials/analysis tools. N.A.A.-H. and N.A.H. analyzed the data. N.A.H. and N.A.A-.H. wrote the article. Finally, all authors reviewed the article.

Ethics Approval

This study was conducted in compliance with the Declaration of Helsinki, and the Ethics Committee at Faculty of Medicine, Beni-Suef University provided its approval (FMBSUREC/02102022/Abd-elhameed).

Informed Consent

Written informed consent was obtained from all subjects before the study.

Author Disclosure Statement

No competing financial interests exist.

Funding Information

No funding was received for this article.

References

Alper

, Zhang

, Pashankar

. Correlation of erythrocyte sedimentation rate and C-reactive protein with pediatric inflammatory bowel disease activity. J Pediatr Gastroenterol Nutr, 2017; 65(2):e25–e27.

Ayeldeen

, Shaker

, Khairy

, et al. Signature of micro RNA 146a/215 and IL-6/TGF-β levels in a cross-link axis between obesity and colorectal cancer. Noncoding RNA Res, 2023; 8(2023):187–191; https://doi.org/10.1016/j.ncrna.2023.01.001

Baumgart

, Carding

. Inflammatory bowel disease: Cause and immunobiology. Lancet, 2007; 369(9573):1627–1640.

Baumgart

, Sandborn

. Crohn's disease. Lancet, 2012; 380:1590–1605.

Chen

, Xia

, Deng

, et al. Emerging microRNA biomarkers for colorectal cancer diagnosis and prognosis. Open Biol, 2019; 9(1):180212.

Chen

, Liu

, Zhao

, et al. Plasma long noncoding RNA expression profile identified by microarray in patients with Crohn's disease. World J Gastroenterol, 2016; 22:4716–4731.

Chen

, Wang

, Feng

, et al. The relationship between C-reactive protein/albumin ratio and disease activity in patients with inflammatory bowel disease. Gastroenterol Res Pract, 2020; 2020:3467419.

Corti

, Fassino

, Marcucci

, et al. Oligometric tumor necrosis factor-? Slowly converts into the reactive forms at bioactive levels. Biochem J, 1992; 284(3):905–910.

Dalal

, Chang

. The microbial basis of inflammatory bowel diseases. J Clin Invest, 2014; 124(10):4190–4196.

10.

Deldar

MAP

, Mirzaei

, Ashrafizadeh

, et al. Wnt/β-catenin signaling as a driver of hepatocellular carcinoma progression: An emphasis on molecular pathways. J Hepatocell Carcinoma, 2021; 8:1415–1444.

11.

Dubois-Camacho

, Diaz-Jimenez

, De la Fuente

, et al. Inhibition of miR-378a-3p by inflammation enhances IL-33 levels: A novel mechanism of alarming modulation in ulcerative colitis. Front Immunol, 2019; 10:2449.

12.

Duttagupta

, DiRienzo

, Jiang

, et al. Genome-wide maps of circulating miRNA biomarkers for ulcerative colitis. PLoS One, 2012; 7:e31241.

13.

Eltayeb

, Aljafari

, Algazy

, et al. Role of tumor necrosis factor alpha α and interleukin 10 gene polymorphisms in irritable bowel syndrome patients. Int J Health Sci Res, 2017; 7:100–106.

14.

Eltayeb

, Shams

, Babikr

, et al. Serum levels of interferon gamma INF-γ and interleukin 10 Il-10: An immunological aspect among irritable bowel syndrome patients. Pak J Biol Sci, 2020; 23:898–903.

15.

, Wu

, Liu

. C-reactive protein and cardiovascular disease: From animal studies to the clinic. Exp Ther Med, 2020; 20(2):1211–1219.

16.

Han

, Zhang

. Role of long non-coding RNA LINC00641 in cancer. Front Oncol, 2022; 11:829137.

17.

Hart

, Rajab

, Alebraheem

, et al. C-reactive protein and cancer—Diagnostic and therapeutic insights. Front Immunol, 2020; 11:595835.

18.

, Su

, Lei

, et al. LINC00641/miR-582-5p mediate oxaliplatin resistance by activating autophagy in gastric adenocarcinoma. Sci Rep, 2020; 10:14981.

19.

Jarczak

, Nierhaus

. Cytokine storm—Definition, causes, and implications. Int J Mol Sci, 2022; 23(19):11740.

20.

Jeong

, Baek

, Kim

, et al. C-reactive protein level as a marker for dyslipidemia, diabetes and metabolic syndrome: Results from the Korea National Health and Nutrition Examination Survey. BMJ Open, 2019; 9:e029861.

21.

Kaplan

, Windsor

. The four epidemiological stages in the global evolution of inflammatory bowel disease. Nat Rev Gastroenterol Hepatol, 2021; 18:56–66.

22.

Karoui

, Laz

, Serghini

, et al. Correlation of C-reactive protein with clinical and endoscopic activity in patients with ulcerative colitis. Dig Dis Sci, 2011; 56:1801–1805.

23.

Khaliefa

, Desouky

, Hozayen

, et al. miRNA-1246, HOTAIR, and IL-39 signature as potential diagnostic biomarkers in breast cancer. Noncoding RNA Res, 2023; 8(2023):205–210; https://doi.org/10.1016/j.ncrna.2023.02.002

24.

Khalil

, Shaker

, Hasona

. lncRNA H-19 and miR-200a implication and frequency of lncRNA H-19 rs2170425 SNP in ulcerative colitis and Crohn's disease. Comparat Clin Pathol, 2023; 32:565–571; https://doi.org/10.1007/s00580-023-03465-2

25.

Khan

, Patel

, Shah

, et al. Albumin as a prognostic marker for ulcerative colitis. World J Gastroenterol, 2017; 23(45):8008–8016.

26.

Lamb

, Kennedy

, Raine

, et al. British Society of Gastroenterology consensus guidelines on the management of inflammatory bowel disease in adults. Gut, 2019; 68:s1–s106.

27.

Levitt

, Levitt

. Human serum albumin homeostasis: A new look at the roles of synthesis, catabolism, renal and gastrointestinal excretion, and the clinical value of serum albumin measurements. Int J Gen Med, 2016; 9:229–255.

28.

Livak

, Schmittgen

. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−delta delta c(t)) method. Methods, 2001; 25(4):402–408.

29.

Matsuoka

, Kobayashi

, Ueno

, et al. Evidence-based clinical practice guidelines for inflammatory bowel disease. J Gastroenterol, 2018; 53(3):305–353.

30.

Nakase

, Sato

, Mizuno

, et al. The influence of cytokines on the complex pathology of ulcerative colitis. Autoimmun Rev, 2022; 21(3):103017.

31.

O'Brien

, Hayder

, Zayed

, et al. Overview of microRNA biogenesis, mechanisms of actions, and circulation. Front Endocrinol, 2018; 9:402.

32.

Padua

, Mahurkar-Joshi

, Law

, et al. A long noncoding RNA signature for ulcerative colitis identifies IFNG-AS1 as an enhancer of inflammation. Am J Physiol Gastrointest Liver Physiol, 2016; 311:G446–G457.

33.

Sayar

, Kurbuz

, Kahraman

, et al. A practical marker to determining acute severe ulcerative colitis: CRP/albumin ratio. North Clin Istanb, 2020; 7(1):49–55.

34.

, Shen

, Zheng

, et al. The role and mechanisms of action of microRNAs in cancer drug resistance. Clin Epigenet, 2019; 11:25.

35.

Solem

, Loftus

, Tremaine

, et al. Correlation of C-reactive protein with clinical, endoscopic, histologic, and radiographic activity in inflammatory bowel disease. Inflamm Bowel Dis, 2005; 11(8):707–712.

36.

Sproston

, Ashworth

. Role of C-reactive protein at sites of inflammation and infection. Front Immunol, 2018; 13(9):754.

37.

Statello

, Guo

, Chen

, et al. Gene regulation by long non-coding RNAs and its biological functions. Nat Rev Mol Cell Biol, 2021; 22:96–118.

38.

Vespa

, D'Amico

, Sollai

, et al. Histological scores in patients with inflammatory bowel diseases: The state of the art. J Clin Med, 2022; 11:939.

39.

Wang

, Zhang

, Yu

, et al. Circulating microRNA223 is a new biomarker for inflammatory bowel disease. Medicine (Baltimore), 2016; 95:e2703.

40.

Woźniak

, Barańska

, Małecka-Panas

, et al. The prevalence, characteristics, and determinants of anaemia in newly diagnosed patients with inflammatory bowel disease. Gastroenterol Rev, 2019; 14(1):39–47.

41.

, Yu

, Li

, et al. Wnt/β-catenin signaling in cancers and targeted therapies. Sig Transduct Target Ther, 2021; 6:307.