Transcriptome Analysis of mRNA in Uterine Leiomyoma Using Next-generation RNA Sequencing

Author(s):

Shadab Anjum, Tahreem Sahar, Aruna Nigam and Saima Wajid*   Pages 1703 - 1718 ( 16 )

Abstract:

<P>Background: Uterine leiomyoma is a benign smooth muscle tumor of monoclonal nature in the female reproductive tract and is one of the major health problems. More than 70% of the female population suffers from uterine leiomyoma in their lifetime and in the advanced condition, it is associated with pregnancy complications and infertility. </P><P> Objective: Characterization and relative expression of mRNA transcripts through transcriptome profiling in uterine leiomyoma and adjacent normal myometrium. </P><P> Methods: Uterine leiomyoma tissue of an Indian female, age 32 years, with a family history of leiomyoma (evident from mother’s hysterectomy for the same pathology) was used. Patient showed 9 multiple large lesions appearing heterogeneously, deforming the uterine contour and causing distortion and splaying of the endometrial cavity showing disease aggressiveness was taken for Next-generation sequencing (NGS) to develop whole transcriptome profile along with the adjacent normal myometrium as control. The validation of the relative expression of the selective transcripts was done using Real-Time PCR. </P><P> Results: The transcriptome profile indicated 128 genes up-regulated and 98 down-regulated, with the Log2 fold change ≥ 2 and P ≤ 0.05, highlighting the molecular network closely associated with focal adhesion, hyaluronan and MAPK-signaling pathways. The mean relative fold change obtained from quantitative PCR as well as the P-values of 10 selected transcripts evaluated from student’s t-test were as follows: BCAN: 7.93 fold (p-value =0.0013); AAK1: 2.2 fold (p-value =0.0036); PCBP3: 3.4 fold (p-value =0.0197); MOV10L1: 3.4 fold (p-value =0.0062); TWISTNB: 1.8 fold (p-value =0.006); TMSB15A: 2.1 fold (p-value =0.0023); SMAD1: 0.8 fold (p-value =0.0112); ANXA1: 0.6 fold (p-value =0.0012); FOS: 0.6 fold (p-value =0.0191); SLFN11: 0.56 fold (p-value =0.0001). </P><P> Conclusion: The present study provides a roadmap, towards the analysis of genes and their roles in corresponding pathways throwing light on their possible involvement in the pathology of the disease.</P>

Keywords:

Uterine leiomyoma, fibroid, transcriptome, next-generation sequencing, genes, real-time PCR.

Affiliation:

Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110062, Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110062, Department of Obstetrics and Gynecology, HIMSR and HAH Centenary Hospital, Jamia Hamdard, New Delhi, 110062, Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110062

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