Transfer Of Bacterial Plasmid And Ectopic Expression Of MAGE-A4 Gene In ESCC Cell Line (KYSE-30) And EvaluateTumoral Behavior

Publish Year: 1395
نوع سند: مقاله کنفرانسی
زبان: English
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NASTARANCANSER02_055

تاریخ نمایه سازی: 22 دی 1396

Abstract:

Esophageal cancer is responsible for the death of over ten million people worldwide annually. Thisis the eighth most common cancer in the world, the sixth cancer leading to death and the third mostcommon gastrointestinal malignancy. There are several treatment methods for cancer; many typesof cancers are associated with the reactivation of genes in the germ and fetal layer.One of the mostimportant categories of these genes isCancer Testis (CT) of genes or CG genes and also one of themost important CTAs is related to the MAGE family. In this study, the behavioral changes of theesophageal cancer cell after receiving MAGE-A4 gene have been investigated regarding the invasionand proliferation. Transferring the plasmid containing Mage-a4 gene to ecoli-top 10 bacteria toproliferate. Isolation of plasmid and double enzyme digestion to confirm the desired piece and thencultivating two cell lines of hek293 and kyse30 to transfer plasmid throughlipofectionto thementioned cell lines.Conducting Mtt and scratch assay tests, rna extraction and then synthesizingcDNA for real time pcr. A 6-unit increase of the expression of mage-a4 gene in both cell lines of kyse-30 and hek293. Mtt test results are as follows, KYSE-30; OD untransfect (3.73) OD transfect (2.78)and HEK-293; OD untransfect (1.159) OD transfect (1.153). The results of the scratch assay testindicated that tumor cell line to which the MAGE-A4 gene was transferred has less invasion. In MTTassay test, the mage-a4 gene transfer to hek-293 cell linecaused no change in transfected andUntransfectedcell proliferation. But in the tumor cell line (kyse-30) we witnessed a decrease in cellproliferation in cell lines to which MAGE-A4 gene had been transfected. The results of the scratchassay test indicated that the cell invasion rate was reduced inkyse-30 cell line to which the mage-a4gene was transfected.

Authors

Maryam Ashkar

Department Of Microbial Biotechnology, Islamic Azad University Of Damghan, Iran