mohammadkazem heydari

CD44, a cell-surface receptor and a key player in cellular signaling, can act as both tumor suppressor and promoter. This study aimed to investigate the association of CD44 rs13347C>T variants with prostate neoplasms, including both benign prostatic hyperplasia (BPH) and prostate cancers using a case-control and bioinformatics approach. Genomic DNA was extracted from 545 blood samples (225 BPH, 225 prostate cancers, and 95 control) and the CD44 rs13347C>T genotypes were identified using PCR-RFLP. We explored miRNA interactions using the miRNASNP-v3 database and GeneMANIA for co-expression networks. Results showed cancer patients had significantly higher PSA levels compared to both controls (p= 0.03) and BPH (p= 0.01). Additionally, digital rectal examination-positive and smoker BPH patients showed significantly the increased cancer risk (p= 0.004, p= 0.046). Prostate cancer group indicated significantly higher frequency of CD44 rs13347C>T mutant allele compared to control and BPH groups, particularly in TT and CT+TT genotypes (p < 0.05). miRNA SNP-v3 database predicted the mutant allele of CD44 rs13347C>T could lose 1 and gain 6 miRNAs for a new site created. Co-expression analysis revealed a direct interaction between CD44 and aryl hydrocarbon receptor (AHR), a gene known to be dysregulated in smokers. Furthermore, these genes alone display co-expression interactions with integrin subunit alpha 4 (ITGA4), protein plays a paradoxical role, both suppressing and promoting tumors. Based on the findings, the mutant allele of CD44 rs13347C>T may disrupt miRNA binding, which may potentially impact CD44, AHR, and ITGA4 expression in smokers, possibly contributing to prostate cancer progression.

Today, numerous antimicrobial and anticancer properties have been reported for plant lectins due to theirability to bind to carbohydrates. The Urtica dioica agglutinin (UDA lectin) is a monomeric, small, and low molecularweight glycoprotein. It has attracted the attention of many researchers for identification, treatment, and other clinicalpurposes.

The aim of this study is the optimization of the chitin affinity chromatography based on Sepharose 4B (CNBractivated Sepharose 4B) for the rapid purification of UDA lectin from Urtica dioica rhizome.

The chitin ligands were dissolved in 40% Trichloroacetic acid and attached to Sepharose4B according to the Amersham-Biosciences instructions. The attachment of the ligand to the Sepharose 4B beads wasinvestigated by Fourier transform infrared (FTIR) spectroscopy. An acidic crude extract of nettle rhizome passes fromchromatographic columns in two sizes with dimensions: 24 x 0.51 cm and 8.44 x 0.86 cm. Quantity and quality of purifiedlectin were calculated by the Bradford microplate method SDS-PAGE gel electrophoresis and human erythrocyte cell(RBC) hemagglutination, respectively.

The analysis of FTIR spectrograms showed that major changes were observed in the fingerprint regions. Besides,due to the dissolution of Sepharose 4B and chitin in the aqueous phase, this difference was not significant in the Imineand Nitrile regions. On the other hand, the comparative results of purification chromatograms showed that increasingthe column length causes a smaller half-width and increases the length of the purified peak. Also, it leads to high-qualitypurified UDA lectin, with a molecular weight of almost 12.5 kDa in gel electrophoresis. Hemagglutination activityon trypsinized red blood cells was displayed, and agglutination of purified UDA lectin started at least at 300 μg.mL-1concentration.

According to our findings, we suggested that dissolving chitin in the polar solvent of Trichloroacetic acid,using Sepharose 4B as the beads of a matrix, and increasing the column length might lead to a decrease in the half-widthof the peak. These can increase the purity and concentration of purified UDA lectin, and speed up the purification process. These findings could be used by researchers to accelerate the purification of UDA lectin in other studies, dealing with drug delivery systems, ELISA techniques, and cell growth.

Recurrent spontaneous abortion (RSA) is one of the causes of infertility following fetus creation. This can lead to the reduction of the fertility rate in various populations. Collagens’ structure, their ratios to each other, and their connections to various receptors are some of the key players in successful fetus implantation. Among them, COL12A1 has a special role because of its very high expression in the uterus. Also, the CD44 protein as a cell adhesion molecule which is a receptor for some of the collagens plays a significant role in RSA. The aim of the study is to investigate the association of CD44-rs13347 and CoL12A1-rs970547 with RSA in a case-control base, and the impacts of COL12A1-rs970547 on protein structure. To genotype single nucleotide polymorphisms (SNPs) of the genes, the PCR-RFLP method was performed on the 124 RSA and 124 control samples. The results were analyzed by the binary-logistic regression method (p-value≤0.05). The SNPs’ effect on the proteins’ structure was analyzed by PSIPRED, HOPE, LOMET, and chimera-USF. Proteins signaling pathway and physical interaction between COL12A1 and CD44 were investigated by KEGG-pathway and GeneMANIA, respectively. Results showed that TT (P= 0.032) genotype of CD44-rs13347 increased the risk of RSA while the CT (P= 0.027) genotype of CD44-rs13347, TT (P= 0.044) genotype, and T (P= 0.019) mutant allele of COL12A1-rs970547 decreased the risk of RSA. Moreover, 3D structures investigation indicated that COL12A1-rs970547 may affect the structure of COL12A1 and its interaction with Integrins. The analysis of the signaling pathway and proteins’ physical interaction network also revealed the interaction of COL12A1 and CD44 with MMP2 and MMP9. On this base, we recommend that T allele of COL12A1-rs970547 has a protective feature against RSA, especially in homozygous form by improving their interaction with Integrins and probably MMPs, too. On the other hand, the CD44-rs13347 probably has an indirect influence on the attachment of the fetus to the extracellular matrix by affecting the MMPs and finally leading to a greater risk of RSA.

Expression of CD44 variant 6 (CD44v6) as a homing-associated cell adhesion molecule (HCAM),hasproved to change most cancer cells. Aim of the study is the effect of mutant allele of CD44 (rs8193C>T),Pum2regulatory element as a prognosis factor of prostate neoplasms: a case-control,in silico studies in the Mazandaranprovince-Iran.

In a case-control study,CD44-rs8193C>T genotyping of the 420 prostate neoplasms (210benign prostatic hyperplasia (BPH) patients,210 prostate cancer patients),150 healthy samples are performedby the touchdown polymerase chain reaction with confronting two-pair primers (PCR-CTPP) method. The T mutantallele effects on the mRNA structure,cell pathways were also investigated in silico methods.

Our results showed that the increase of T mutant allele frequency was significantly associated with BPHcompared with prostate cancer. Furthermore,results showed TT genotype was significantly associated with BPH[odds ratio (OR),0.572,P,0.015],and also influenced the CD44v6 transcript secondary structure,miRNA binding,and regulatory element-binding site for Pum2 protein. Attachment of Pum2 to standard CD44 transcript may lead totranscript isoform-switching,shift-expression to a variety of CD44 isoforms,which can trigger some of the cellsignaling pathways,such as Nanog-Stat,PKC-Nanog,and PKC-Twist.

Based on this,the presence of the T mutant allele of CD44 (rs8193C>T) in the populations may createa regulatory element-binding site for Pum2. So,it could be known as a prognosis factor,prediction of prostateneoplasms. However,more comprehensive studies in different populations (with various ethnicities,large populationsizes),and also CD44v6 gene expression studies in protein,transcript levels are required to confirm our data.