MicroRNA Levels in Cervical Cancer Samples and Relationship with Lesion Grade and HPV Infection

Abstract

Background: miR-21, miR-214, and miR-let-7a are three validated and well-known miRNAs. miR-21 is described as an “oncomir,” while miR-214 and miR-let-7a are described mainly as tumor suppressors. The role of these miRNAs remains unclear in cervical cancer, an important malignancy among women worldwide and responsible for many deaths every year. Objective: The objective of this study was to describe the expression profile of miR-21, miR-214, and miR-let-7a in plasma and cervical scraping from a control group and patients with different grades of cervical lesions and invasive cervical cancer, and then correlate with HPV infection groups. Methods: Plasma and cervical scraping were submitted to DNA and RNA extraction. HPV detection and typing were performed by conventional PCR followed by PAGE to amplicons interpretation. The miRNA relative expression in plasma and cervical scraping samples was performed by real-time PCR using specific TaqMan probes. Results: miR-21 (p=0.0277) and miR-214 (p=0.0151) were up-regulated in cervical scraping samples of the invasive cervical cancer (ICC) group. However, miR-214 was also up-regulated in the LSIL group (p=0.0062). Both miRNAs were not related to HPV infection. However, miR-let-7a was higher in HPV positive plasma samples (p=0.0433) than in HPV negative plasma samples, and the correlation analysis confirmed the association between the levels of this miRNA with the presence of HPV (p=0.0407; r=0.3029), but not with lesion grade (p>0.05). Conclusion: Our results suggest that miR-21 is related to cervical cancer progression and miR-214 appears to have an ambiguous role in cervical lesions. miR-let-7a may be upregulated at the systemic level in patients with HPV infection.

Systematic review of circulating MICRORNAS as biomarkers of cervical carcinogenesis

“…The mean age was 39.81 years (ranging from 27.5 to 50 years), but six studies did not report the age of included donors [ 11 ] [ 8 ] [ 13 ] [ 15 ] [ 16 ] [ 23 ]. Still, in relation to the control group, seven studies reported that samples were tested for HPV infection [ 14 ] [ 17 ] [ 18 ] [ 19 ] [ 23 ] [ 24 ] [ 25 ], and one study declared that the samples were not tested [ 11 ] and other studies did not report any information on the assessment of HPV infection. Among the studies in which samples were tested for HPV infection, one reported using the liquid-cytology method [ 14 ], and four reports of HPV infection were evaluated by PCR [ 17 ] [ 23 ] [ 24 ] [ 25 ], identifying the presence of HPV types 16, 66 and 68.…”

“…Still, in relation to the control group, seven studies reported that samples were tested for HPV infection [ 14 ] [ 17 ] [ 18 ] [ 19 ] [ 23 ] [ 24 ] [ 25 ], and one study declared that the samples were not tested [ 11 ] and other studies did not report any information on the assessment of HPV infection. Among the studies in which samples were tested for HPV infection, one reported using the liquid-cytology method [ 14 ], and four reports of HPV infection were evaluated by PCR [ 17 ] [ 23 ] [ 24 ] [ 25 ], identifying the presence of HPV types 16, 66 and 68. Two studies reported that HPV infection was not detected in women in the control group [ 18 ] [ 19 ], without mentioning the method used for HPV detection.…”

“…The mean age among studies that reported this information was 38.2 years (ranging from 32 to 50). Six studies reported that CIN samples were tested for HPV infection [ 14 ] [ 17 ] [ 19 ] [ 23 ] [ 24 ] [ 25 ] and the others did not report any information about HPV infection in this patient group. Kong et al [ 14 ] reported HPV infection in all patients included in the CIN group and the same was reported by Farzanehpour et al [ 17 ], who also reported that HPV types 16 and 53 were identified.…”

“… miR-21-5p miR-199a-5p miR-155-5p miR-34a-5p miR-145-5p miR-218-5p No miR-21-5p: 0,97 miR-199a-5p: 0,831 miR-155-5p: 0,719 miR-34a-5p: 0,888 miR-145-5p: 0,575 miR-218-5p: 0,609 Combined: 0,942 miR-21-5p miR-199a-5p miR-155-5p Combined: 0,969 miR-34a-5p miR-145-5p miR-218-5p Combined: 1 miR-21-5p miR-199a-5p miR-155-5p miR-34a-5p miR-145-5p miR-218-5p miR-21-5p: 88 miR-199a-5p: 86.7 miR-155-5p: 93.8 miR-34a-5p: 89.1 miR-145-5p: 67.7 miR-218-5p: 83.3 Combined: 100 miR-21-5p miR-199a-5p miR-155-5p Combined: 100 miR-34a-5p miR-145-5p miR-218-5p Combined: 100 miR-21-5p miR-199a-5p miR-155-5p miR-34a-5p miR-145-5p miR-218-5p miR-21-5p: 98 miR-199a-5p: 91.7 miR-155-5p: 88.3 miR-34a-5p: 91.7 miR-145-5p: 60 miR-218-5p: 72.7 Combined: 78.9 miR-21-5p miR-199a-5p miR-155-5p Combined: 92.8 miR-34a-5p miR-145-5p miR-218-5p Combined: 100 miR-21-5p miR-199a-5p miR-155-5p miR-34a-5p miR-145-5p miR-218-5p Significant up-regulation of miR-21-5p, miR-199a-5p and miR-155-5p and down-regulation of miR-145-5p, miR-34a-5p and miR-218-5p was observed in women with LSIL/HSIL and CC when compared to healthy controls. Hoelzle et al [ 24 ] 2021 qPCR miR-23a-3p 2 −ΔΔCt Higher plasmatic miR-let-7a expression levels awithociated with HPV infection miR-21 miR-let-7a miR-214 Yes N/I N/I N/I No significant differences were found between groups. Ocadiz-Delgado et al [ 25 ] 2021 qPCR Beta2mHu 2 −ΔΔCt N/I miR-15b miR-34a miR-218 No LSIL miR-34a: 0,9952 miR-15b: 0,6765 miR-218: 0,9872 CC miR-34a: 0,9505 miR-15b: 0,8173 miR-218: 0,8421 N/I N/I MiR-15b expression was higher in LSIL group and even higher in CC patients compared to healthy controls.…”

Disease progression role as well as the diagnostic and prognostic value of microRNA-21 in patients with cervical cancer: A systematic review and meta-analysis

“…Many studies have most consistently revealed the upregulated expression of microRNA-21 in cervical carcinogenesis process [ 41 , 73 , 98 , 99 , 111 – 113 ]. It has quite a complex role in cancer development and tumorigenesis with many targets in which a variety of mechanisms play a role.…”