CLEC4E as a New Tumor Marker for Early Diagnosis of GIT Tumors


Rasha Hasan Jasim, Hadeer Haider Abdualameer

Department of Chemistry, Faculty of Education for Girls, University of Kufa, Iraq.

*Corresponding Author E-mail:



During the period from the beginning of December 2018 to the end of May 2019, 108 resident of Babylon were included to participate in the current study, were divided into two main groups: 81 patients with GIT diseases, their age ranged from 12-80 years; and the controls group (27 individuals) with the age 17-71 years old. Furthermore, the patient group was divided into two subgroups, the first involved 51 patients with GIT tumors and the other with non-tumoral diseases of GIT (30 individuals suffered non tumoral GIT illness were enrolled as a pathological control group). CLEC4L, Serotonin were evaluated in the sera samples of the study groups. The current study revealed statistical significant increase in the levels of serotonin while it showed significant decrease in the levels of CLEC4L in the sera testers of patients with malignant GIT tumors when compared to benign GIT tumors, pathological GIT control, and healthy control groups. The present study showed that the relationship between CLEC4E with serotonin became negative. Serotonin showed the (88%) sensitivity among the assessed criteria followed by CLEC4L (76%).


KEYWORDS: GIT, Cancers, Tumors, Serotonin, Glycoprotein.




Lectins stand for proteins of non-immune origin with widespread natural distribution biologically, which distinguish and reversibly combine to carbohydrates and glycoconjugates. They have been free or interconnected to cell surfaces via particular binding positions1. Lectins vary in amino acid structure, the metals association, 3D arrangement and molecular weight2. Lectins are found in each organisms class, comprising prokaryotes, plants, and animals3. Lectins were first described in 1888 by Still mark, who observed that crude extracts of castor beans (Ricinus communis) contained a toxic substance named "Ricin" was capable of agglutinating red blood cells and he termed it as phytohemagglutinnins4.


Lectins and their specific properties, principally attributable to their capability for binding glycoconjugates, as significant tools in studies related to numerous scopes of biological, clinical and chemical disciplines3.


Lectins possess various special effects on cells, like agglutination, mitogenic stimulation, rearrangement of cell surface constituents, adapting the action of membrane enzymes, inhibiting bacteriological and fungous development, cell aggregation, toxicity, immunomodulation, among others5.


In a time once biotechnology has speedily bettering, scholars have perceived that a plethora of natural sources to be prospective panaceas in contradiction of malignancy6. These proteins may have dissimilar tasks like biological regulating, defense in contradiction of microorganisms attack, protein storage, carbohydrate transference, mitogenic stimulation, recognizing nitrogen-fixing bacteria of Rhizobium genus, and others7. Numerous lectins fight digestion, survive gut passageway, and have binding for gastrointestinal cells and/or access circulating intact, the plant lectins role in cancer treatment upholding full organic activity comprising particular agglutination of lymphocytes, erythrocytes, spermatozoa, platelets, bacteria and tumor cells, induction of mitosis or cytotoxic effects on lymphocytes8. Binding among surface cell molecules and lectins or internalization into cells include a widespread diversity of signals that are imperative for cell regulating, having9.


·       Down regulating of telomerase activity and angiogenesis inhibition.

·       Increasing drug sensitivity of growth cells, henceforth their utility in designing immunotoxins for cancer treatment.

·       Direct impacts on the immune system by varying the creation of numerous interleukins, or by stimulating definite protein kinases.

·       Lectins Ingestion correspondingly sequesters the existing body pool of polyamines, thus thwarting cancer cell growth.

·       Several lectins has feasibility of binding to ribosomes, constraining protein synthesis10.


Serotonin (5-hydroxytryptamine, 5-HT) has been initially recognized as a vasoconstrictor substance that is released from platelets throughout a blood coagulation, and then as the monoamine neurotransmitter in the brain11. About 95% of serotonin is found in the gut whereas the remaining ratio is found in the brain12. In the central nervous system (CNS), where it has various functions, these include the regulation of mood, appetite, and sleep13. Taking Serotonin orally doesn’t pass into the serotonergic alleyways of the fundamental nervous system, for the reason that it doesn’t cross the blood–brain barrier. However, new evidence recommends that serotonin influence cross endothelial cells of the blood brain barrier (BBB) by means of serotonin transporter14. It serves as a vasoconstrictor and helps to regulate hemostasis and blood clotting15. Furthermore, serotonin can be as well in plants (walnuts, bananas, hickories, tomatoes, pineapples), mushrooms, octopuses, and in the insects poison (scorpions, spiders, wasps)16.


Serotonin has an consequence on the amount of physiologic and interactive function; it shows a very significant roles in normal brain task, gastrointestinal, endocrine, and vascular, and many others17. Serotonin shows a development stimulatory effect on some categories of carcinoma, carcinoid and other tumor cells18. It is essentially employed in oncology as a tumor indicator of gastrointestinal carcinoid, hepatic carcinoid; additionally, serotonin is employed as a particular tumor indicator for gastrointestinal growths of the pancreatic islet cells and intestinal tract19.



Throughout the beginning of December 2018 to the end of May 2019, 108 resident of Babylon were included to participate in the current study, were divided into two main groups: 81 patients with GIT diseases, and 27 individuals from the controls group. Furthermore, the patient group was divided into two subgroups, the first involved 51 patients with GIT tumors and the other with non-tumoral diseases of GIT (30 individuals suffered non-tumoral GIT illness were enrolled as a pathological control group). Samples of GIT cancers were collected from patients registered for treatment at Babel Cancer Center in Babylon. While the samples of benign GIT tumors and non-tumoral diseases of the digestive system were collected from public hospitals (Morgan Hospital, Al-Imam Al-Sadiq Hospital) in Babylon. Cancerous GIT patients group were classified into five subgroups (Esophageal, gastric, hepatocellular, pancreatic, and colorectal carcinoma) according to the primary site of the tumor.


Five milliliters of intravenous blood samples were collected from patients and healthy subjects. The samples were left to coagulate at laboratory temperature, then separated using a centrifuge at 5,000xg for 5 minutes. Sandwich-Enzyme-Linked Immune Sorbent Assay (Sandwich-ELISA) method was applied to evaluate lectin, and serotonin concentrations.



3.1: Evaluation of Lectin Concentration in the Serum Samples of GIT Cancer Patients (at Diagnosis), Benign GIT Tumors Patients, Pathological GIT Controls, and Healthy Control Groups:

Estimation of the CLEC4E in the study groups exposed a statistical significant decrease (p = 0.031, p = 0.008, p = 0.001) in the lectin concentration of malignant GIT samples comparison to benign GIT tumors, pathological GIT controls, and healthful entities; respectively. Highly significant variation was observed when benign GIT tumors and healthy individuals (p = 0.009) groups compared together, while no such results were observed when pathological GIT controls were compared with both benign GIT tumors and healthful individuals collections, as shown in Table 1.


Table 1: Levels of CLEC4L (Mean ± S.D.) Concentration (pg/ml) in the Sera of Tumor Patients and Controls Subjects

Study Groups (n)

CLEC4L Conc. (pg/ml) Mean ± S.D.





582.151 ± 279.553

143 – 1080


Malignant vs Benign


Malignant vs PGITC


Malignant vs Healthy


Benign vs Pathological


Benign vs Healthy


Pathological vs Healthy



669.172 ± 320.079

73 - 1039



776.777 ± 205.24

222- 1148



901.568 ± 245.827

476 – 1270



In order to investigate possible differences in the levels of lecithin as a result of the difference in the sex of the individuals participating in the study, it was divided into implicit subgroups based on their gender. When the comparisons between the different implicit subgroups of the same sex were done, the study showed the presence of a significant decrease (p = 0.041, p = 0.020, p = 0.001) in the levels of evaluated lectin (CLEC4L) in males with cancerous digestive system tumors at what time compared with those with benign digestive system tumors and the subgroup of males with non-tumoral gastrointestinal disorders as well as healthy males. Whereas, the statistical results lacked significant when performing the comparison using ANOVA test among female subgroups except for the women with malignant GIT tumors compared to women with non-tumoral diseases of the digestive system, when the results shown a significant decrease in CLEC4L levels (p = 0.042) of the cancerous female subgroup comparison to the pathological control females.


Gastrointestinal tumor samples participating in the present study were distinguished for patients in the early diagnostic stages and that all samples were not yet subject to any therapeutic interventions, whether surgical, chemical, or radiological. The current study is the first in the investigation of the CLEC4L levels and because it is C-type lectin, the current study is based on the hypothesis of the possibility of its participation in the immune defense anti-inflammatory system, similarly to the role of Mannose Binding Lectin (MBL) whose low level is a direct cause in reviewing susceptibility defensive body against various diseases.


Decrease levels of CLEC4L lectins in malignant tumor specimens may be explained through several hypotheses:

·       Based on the hypothesis that CLEC4L may play an important role in the body's immune response20. Therefore, the decrease in the rates of this protein may be due to an exacerbation of the inflammatory condition caused by the progression of the cancerous injury and decreased defensibility21.

·       The probable reason for the low CLEC4L levels may be due to mutations in the gene responsible for the CLEC4L expression which caused by cancerous shifts that can reduce the production of the CLEC4L subunit22.

·       The study assumes that genetic and environmental factors are concerned in developing CLEC4Ldeficiency and susceptibility to cancerous infection23.

·       Common lectin scholars have concentrated on adopting lectins from dissimilar sources (other than human) in humanoid medicinal fields24. Initial results propose that several lectins, can identify changes of malignant cells and decrease the cancer cell tumorigenicity, therefore may have aids for the immune status for patients25.


Previous study has estimated lectin levels in sera of normal individuals and patients of metastatic breast, ovarian, lung, melanoma cancers, and non-Hodgkin’s lymphoma, and gastrointestinal9,26-29.


3.2: Evaluation of Serotonin Concentration in the Serum Samples of GIT Cancer Patients (at Diagnosis), Benign GIT Tumors Patients, Pathological GIT Controls, and Healthy Control Groups:

Estimation of the serotonin in the study groups revealed a statistical significant increase (p = 0.031, p = 0.035, p = 0.005) of this hormone concentration in the samples of malignant GIT tumors when compared to benign GIT tumors, pathological GIT controls, and healthy individuals, respectively. On the other side, non-significant variations were observed when benign GIT tumors compared with the pathological GIT controls as well as healthy individuals. With the same manner statistically insignificant differences were illustrated when the two controls groups were compared together, as illustrated in Table 2.


Table 2: Levels of Serotonin (Mean ± S.D.) Concentration (ng / ml) in Sera of Tumors Patients and Controls Subjects

Study Groups


Serotonin Conc.


Mean ± S.D.






226.656 ± 33.832

139.9- 269.5


Malignant vs Benign


Malignant vs Pathological


Malignant vs Healthy


Benign vs Pathological


Benign vs Healthy


Pathological vs Healthy



178.313 ± 49.583

140. 3 - 332.7



175.269 ± 27.290

130.2 - 232.5



156.673 ± 37.685

121.1 - 218


In order to study the effect of sex on serotonin levels in the four main study groups, the individuals were divided according to gender. The ANOVA test shows the absence of statistical significance when comparing the sexes in each of the major groups. On the other hand, the present study demonstrated significant differences when comparing serotonin levels in the sera of patients (male or female) with malignant digestive system tumors with their peers of the same sex in groups of non-cancerous GIT tumors patients or the pathological controls group as well as the healthy group. Whereas no statistically significant differences were observed when the comparison was made by the ANOVA test among the male groups with each other or in the female groups.


Based to the chemical nature of serotonin hormone, it is transmitted using carrier (5-hydroxytreptamine transport protein "5-HTT") is responsible for reuptake of serotonin in serotonergic nervous endings30. Increase of the SERT in the membrane of cancerous organs may represent one of the most acceptable theories to explain an increase in the serotonin levels in the testers of patients with malignant GIT tumors31.


The present result was relatively agreed with other studies such as Rasha and Noor study which assessed serotonin levels in several cancer types. Maximum serotonin concentrations have been noted in the lung cancers, after that bladder cancers, whereas the lowermost serotonin concentrations have noted in group of patients with other cancerous tumors like GIT tumors. Beyond treatment, serotonin levels have reduced in the entire cancerous tumors group, just bladder cancer and lymphoma groups have depicted significant increasing in a serotonin levels as compared to their levels at analysis32. Same results were illustrated in the study that carried out in the samples of patients with colorectal cancers33.


Hejazi's study has explained the roles of neurotransmitters as in serotonin in cancers development, via actions on numerous categories of receptors for breast cancer patients than the healthy people34. Another study referred to several finest means for addressing the effect of serotonin in cancer through inhibiting serotonin receptors and not through an inhibiting serotonin transporter. Serotonin directly impacts on cell growth via binding to its receptors. The serotonin-1 and serotonin-2 receptors are expansively stated in the human breast tumor, prostate cancer, bladder malignance cells, and other.


3.3: Study the Relationship of CLEC4E to Serotonin in the Sera Samples of Different GIT Patients and Healthy Controls Groups

The relationship between selected lectin (CLEC4E) and serotonin levels in the groups of patients with GIT diseases as well as healthy controls group was studied. Figure 1 A shows high significant negatively (r = - 0.896 at p < 0.000) correlation when CLEC4E and serotonin levels in the patients group with malignant GIT tumors were combined together, while no such relationship was noted when the correlations were done in the groups of patients with benign tumors, pathological GIT diseases, and healthy controls, as illustrated in Figure 1 (B, C, and D).


Figure 1: Correlation of CLEC4E and Serotonin Concentrations in Sera Samples of (A): Malignant GIT Tumors, (B): Benign GIT Tumors, (C): Pathological GIT Disorders, and (D): Healthy Individuals


As far as we know, no study has been recorded linking serotonin and any type of lectins in the natural or pathological condition, whether that injury is carcinogenic or non-carcinogenic, so the present study is the first of its kind to link these two criteria.


3.4: Specificity of the Evaluated Parameters in Detection of Benign Gastrointestinal Tract Tumors

Individual and combined sensitivity of the parameters (CLEC4L, Serotonin) were evaluated in the group of malignant GIT tumors as summarized in the Table 3 Serotonin allergic was estimated towards gastrointestinal carcinomas (88%) when 29 out of 33 gastrointestinal carcinomas showed an increase in serotonin levels compared to healthy individuals. Furthermore, the lectin (CLEC4L) that was evaluated in the current work came after serotonin, where 76% of patients with malignant GIT tumors showed low levels of lectin compared to the mean lectin recorded in the control group.


These findings reinforced the objective of the present study to investigate the possibility of using parameters as diagnostic tools for GIT cancers.


Table 3: Individual and Combined Sensitivity of The Evaluated Parameters in Gastrointestinal Tract Cancer Patients Group












Based on the results obtained from the current work, a number of conclusions can be reached, the most important of which are:

·      Individual valuable criteria in the current work can be good diagnostic tools for digestive system tumors even in its early stages.

·      Evaluating the criteria of the current work simultaneously with each other increases the possibility of using them as tools to distinguish the stages of carcinogenesis, which contributes to enhancing the doctor's ability to determine the method of treatment more accurately.

·      The age and genetic factors were not the decisive factors in the GIT cancer occurrence, but they are enhanced malignancy progression.

·      In general, level of serotonin decreases with age.

·      The carcinogenicity progresses and symptoms of change in vital parameters in men appear faster and more clearly compared to women.



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Received on 14.05.2020           Modified on 04.07.2020

Accepted on 12.08.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(6):3167-3172.

DOI: 10.52711/0974-360X.2021.00552