Abstract
Purpose : To determine the expression profile of lncRNA H19 in different cervical cancers, and to decipher its function in the growth and metastasis of cervical cancer. Methods : The analysis LncRNA H19 expression was performed using quantitative real time polymerase chain reaction (qRT-PCR). Cell counting kit 8 (CCK8) assay was used to assess the viability of the cells. The cells were transfected with Si-H19 using Lipofectamine 2000 and the metastasis of cells was determined by cell migration and invasion assay. Immunoblotting was used to evaluate the protein expression. Results : The lncRNA H19 expression was considerably enhanced in cervical cancer cells, and was about 2.6 to 5.3 times more in cervical cancer cells relative to non-cancer cells. Inhibition of lncRNA caused significant reduction in cervical cancer cell growth in a time-dependent manner. In addition while silencing of lncRNA inhibited the metastasis of HeLa cells. Cell migration and invasion was about 26 % in Si-H19 transfected cervical cancer cells, relative to 65 % in Si-NC cervical HeLa cells. Similarly, cell invasion was 45 % in Si-H19 cervical HeLa cells relative to the negative control (Si-NC). Inhibition of HeLa cell metastasis was also concomitant with decline of metalloproteinases (MMP)-2 and 9 expression. Conclusion : lncRNA regulates the growth and metastasis of cervical cancer cells. Thus, IncRNA may be an important therapeutic agent for cervical cancer. Keywords : Cervical cancer, lncRNA, Proliferation, Invasion
Highlights
Cervical cancer is among the most commonly detected cancers and is ranked as the 3rd most common type of cancers in women worldwide.Annually, around 0.5 million women are detected with this type of cancer accounting for about 9 % of all the diagnosed cancer cases annually [1]
LncRNA H19 upregulated in cervical cancer cell lines
The quantitative real time polymerase chain reaction (qRT-PCR) results showed that IncRNA H19 expression was enhanced in cervical cancer cells as against the non-cancerous cell line HNC-PI 15
Summary
Cervical cancer is among the most commonly detected cancers and is ranked as the 3rd most common type of cancers in women worldwide. The expression of lncRNA H19 was investigated in different cervical cancer cell lines. Human cervical cancer (HeLa, CaSki, MARQ, C33A, C4-1 and SW756) and non-cancerous (HNCF-PI 15) cells were obtained from Type Culture Collection of Chinese Academy of Sciences, Shanghai, China and were cultured in RPMI 1640 complete media containing 10% FBS and antibiotics (100 U/mL penicillin and 100 U/mL streptomycin), at 37 °C in a humidified chamber. Total RNA was isolated from the cervical cancer cells with the help of RNeasy RNA Isolation Kit following the guidelines of the manufacturer and cDNA was synthesized from it by employing RevertAid cDNA Synthesis Kit (Fermentas). The Si-NC and Si-H19 HeLa cells that reached the lower surface of the membrane were subjected to staining with crystal violet (CV), and the images of CV-stained cells were taken. Student’s t-test was employed for statistical analysis using GraphPad prim 6 software
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