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Published on: February 9, 2024
miR-494 inhibits cervical cancer cell proliferation through upregulation of SOCS6 expression
Lei Cheng1,2, Beihua Kong1, Ying Zhao2
1Department of Obstetrics and Gynecology, Qi Lu Hospital of Shandong University Jinan, Jinan, Shandong 250012, P.R. China.
Abstract:
It is unclear how microRNA (miR)-494 inhibits the proliferation of cervical cancer cells by altering the expression of SOCS6. Therefore, the present study aimed to investigate the molecular mechanism underlying miR-494 regulation of suppressor of cytokine signaling 6 (SOCS6) in human cervical cancer samples and the human cervical cancer HeLa cell line. The expression of miR-494 was determined using reverse transcription-quantitative polymerase chain reaction. In addition, TargetScan was used to predict miR-494 target genes and the luciferase reporter assay was used to determine whether SOCS6 was a direct target of miR-494. The results of the present study demonstrated that compared with the cervical intraepithelial neoplasia and normal cervical tissues, the miR-494 expression level in cervical cancer samples was significantly decreased (P<0.01). In addition, compared with normal cervical tissue, miR-494 expression level was significantly decreased in cervical intraepithelial lesions (P<0.05). Furthermore, the expression of miR-494 was associated with patients with or without lymph node metastasis, clinical stage and depth of stromal invasion (P<0.01); however, miR-494 expression was not identified to be associated with age, tumor size and menopausal status (P>0.05). Transfection of a miR-494 mimic significantly increased the expression level of miR-494 in HeLa cells (P<0.01), and anti-miR-494 transfection decreased the expression of miR-494 (P<0.01). An MTT proliferation assay and Boyden chamber invasion ability assay revealed that miR-494 mimic transfection significantly inhibited the proliferation, and invasion ability of HeLa cells (P<0.01), whereas anti-miR-494 transfection significantly increased the proliferation and invasion ability (P<0.05). SOCS6 was predicted, using bioinformatics, to be the target gene of miR-494 and this was validated using a luciferase reporter assay. Western blot analysis revealed that transfection of miR-494 significantly increased the expression of SOCS6 in HeLa cells, and transfection of anti-miR-494 significantly decreased the expression of SOCS6. Therefore, the results of the present study demonstrated that miR-494 expression in cervical cancer was significantly decreased. Exhibiting a decreased expression level of miR-494 may result in enhanced proliferative and invasive abilities of HeLa cell, thus contributing to the occurrence, and development of cervical cancer.
Insights
MicroRNA (miR)-494 is significantly decreased in cervical cancer, inhibiting cancer cell proliferation and invasion by upregulating suppressor of cytokine signaling 6 (SOCS6). This finding clarifies the molecular mechanism of miR-494 in cervical cancer development.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Cervical cancer remains a significant global health challenge.
- The precise molecular mechanisms by which microRNAs regulate cancer progression are not fully understood.
- MicroRNA-494 (miR-494) has been implicated in various cancers, but its role in cervical cancer requires further elucidation.
Purpose of the Study:
- To investigate the molecular mechanism of miR-494 in regulating the expression of suppressor of cytokine signaling 6 (SOCS6) in cervical cancer.
- To determine the effect of miR-494 on the proliferation and invasion of human cervical cancer cells.
- To correlate miR-494 expression levels with clinical parameters in cervical cancer patients.
Main Methods:
- Quantitative reverse transcription polymerase chain reaction (RT-qPCR) for miR-494 expression analysis.
- Bioinformatic prediction (TargetScan) and luciferase reporter assays to validate miR-494 targeting of SOCS6.
- In vitro assays including MTT and Boyden chamber assays to assess cell proliferation and invasion.
- Western blot analysis to evaluate SOCS6 protein expression.
Main Results:
- miR-494 expression was significantly decreased in cervical cancer tissues compared to normal and pre-cancerous tissues.
- Decreased miR-494 expression correlated with lymph node metastasis, clinical stage, and stromal invasion depth.
- miR-494 mimic transfection inhibited proliferation and invasion of HeLa cells, while anti-miR-494 transfection enhanced these abilities.
- SOCS6 was confirmed as a direct target of miR-494, and miR-494 upregulation led to increased SOCS6 expression.
Conclusions:
- miR-494 acts as a tumor suppressor in cervical cancer by inhibiting cell proliferation and invasion.
- The tumor-suppressive function of miR-494 is mediated through the upregulation of its direct target, SOCS6.
- Reduced miR-494 expression contributes to cervical cancer progression and may serve as a potential therapeutic target.
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