Evaluation of let7-i inhibition of melanoma cell line metastasis (A-375) (C6181)
Abstract
Tumors can be benign (noncancerous) or malignant (cancerous). Benign tumors are not cancerous. miRNAs are small RNA molecules of 25-21 nucleotides that bind to target mRNA and play a key role in regulating gene expression. These RNAs are classified into two categories: suppressor tumors and oncogenes. One of the tumor suppressor miRNAs that is reduced in melanoma is Let7 i. As a result of reduced expression of this miRNA, the expression of genes is disturbed. These genes are involved in migration and metastasis.
Materials & Methods: For this purpose, first A-375 and C6181 cancer cells were cultured in RPMI culture medium containing 10% FCS at 37 ° C and 5% CO2, then Let7-i gene with vector and transfectant reagent Lipofectamin2000 It is added to the culture medium, incubation is performed, after the required time the expression of Let7i and the studied genes were measured by qRT-PCR. The growth and proliferation of cancer cells were assessed by MTT technique. The rate of migration and metastasis of cancer cells after Let7-i transfection was assessed by Wound healing assay.
Results: Let7i was declining in melanoma cancer cell lines. The expression level of let7i was measured by RT-qPCR and the result showed that the expression level of this microRNA decreased in both melanoma cell lines compared to NHEM as a normal skin cell line. WoundHelling data showed that Let7i reduced A-375 cell migration 24 hours after zero time compared to control cells.