ABSTRACT
Objective To establish a construction method for luciferase-microRNA (miRNA) sponge adenovirus, and to verify the binding efficiency of the expression product of the adenovirus and miRNA. Methods DNA fragments that can be partly complementary to miRNA-126 were chemically synthesized. The miRNA-126 sponge fragments were amplified by PCR, and cloned into pMD-18T vector and the 3'-end non-coding region of psiCheck2 plasmid; hRluc-miRNA-126 sponge was further cloned into Ad-Track plasmid. After linearization by Pme I, Ad-Track-hRluc-miRNA-126 sponge plasmid was transformed into competent Escherichia coli strains BJ5183 to homologous recombinate with Ad-Easy plasmid; recombinant Ad-Easy-hRluc-miRNA-126 sponge plasmid was identified by Pac I, and then transfected into 293 cells to produce adenovirus. Adenovirus was used to infect human umbilical vein endothelial cells (HUVECs), and the effect of miRNA-126 on cell migration ability was detected by scratch test. Results The miRNA-126 sponge had a good complementary base-pairing relationship with miRNA-126; double luciferase experiment showed that miRNA-126 could directly act on hRluc-miRNA-126 sponge. Cloning hRluc-miRNA-126 sponge into Ad-Track plasmid made it possible to synchronously monitor the efficiency of cellular transfection and miRNA inhibition. Ad-Easy-hRluc-miRNA-126 sponge plasmid was successfully established by homologous recombination of Ad-Track and Ad-Easy, identified by Pac I digestion with 4.5 kb fragments. When transfected into 293 cells, luciferase-miRNA sponge adenovirus was obtained 7 days later. Overexpression of miRNA-126 inhibited the expression of hRluc-miRNA-126 sponge (P<0.01). Infection with miRNA-126 sponge adenovirus inhibited the migration of HUVECs (P<0.05). Conclusion Luciferase-miRNA sponge adenovirus has been successfully established. MiRNA-126 has a high binding efficiency with the expressed product of the adenovirus..
ABSTRACT
The mortality rate of lung cancer has been high all over the world. In recent years, targeted drugs have become a new and effective method for the treatment of advanced lung cancer, but the early diagnosis and long-term effective treatment are still great challenges in lung cancer. Circular RNA (circRNA) is a unique RNA molecule with ring structure, which exhibits excellent stability and expression specificity. Increasing studies found that the expression of several circRNAs in tumors were abnormal. This abnormal expression is not only related to malignancy of tumors, but also involved in regulating the progress of tumors, which provides a new way for the diagnosis and treatment of tumors. Therefore, this article reviews the expression, the diagnostic and prognostic value as well as pathogenesis of circRNAs in lung cancer in order to find new targets for early diagnosis and treatment of lung cancer. .
ABSTRACT
Lung cancer is the most commonly diagnosed cancer and the leading cause of cancer death in China. In recent years, therapies for oncogenedrivers and immune checkpoints have proved inspiring. Circular RNA (circRNA), which is a kind of RNA with covalent ring structure relating to stages and metastasis of cancer, has many special biological functions in physiological processes, diseases and so on. Thus, circRNA is expected to be a potential biomarker for cancer prediction and treatment in view of its high conservation and tissue-specific. However, function analysis and regulatory mechanism of circRNA in lung cancer come so far remains unclear and limited literatures are available. In this review, we highlight the research history, formation mechanism, biological function of circRNA and research progress in cancer, especially in lung cancer. We mean to provide theoretical evidences and new ideas for researches on circRNAs in lung cancer.
Subject(s)
Animals , Humans , Lung Neoplasms , Genetics , Metabolism , RNA , GeneticsABSTRACT
Recently, with the development of RNA research techniques, a wide variety of circular RNAs (circRNAs) have been discovered and some of them are confirmed to have crucial biological functions. CircRNAs arise from exons (i.e. exonic circRNAs) or introns (i.e. intronic circRNAs). Acting as microRNA sponges or combining with proteins, circRNAs participate in the regulation of gene expression and influence the activity of some proteins. In addition, some circRNAs even encode proteins. More importantly, several circRNAs play a key role in the occurrence and progression of some tumors, including stomach, liver, colon, breast, cervical, and ovarian cancers. Therefore, circRNAs may be a novel type of diagnostic marker and therapeutic target of cancers.