According to the findings of a study that was published in Scientific Reports, RAB27A knockdown leads to an increase in the proliferation, migration, and invasion of colon cancer cells. This occurs after RAB27A has been reduced by RAB 27A knockdown.
The researchers also discovered that RAB27A encourages expression at an abnormal location.
They claimed that the findings demonstrate that RAB27A is of crucial importance for the development of colon cancer and that it has the potential to be a prognostic indicator in addition to being a potential target for treating colorectal cancer.
According to the findings of the research conducted, colorectal cancer “develops when colonic epithelial cells acquire a sequence of genetic or epigenetic alterations that promote cell growth and survival.”
Researchers believe that the identification of new genes that are connected with the development of colorectal cancer is vital for the early detection and treatment of the disease.
It has been discovered that Rab proteins can operate as oncoproteins in the process of cancer development. For example, the knockdown of RAB10 can block the growth cycle and colony formation in hepatocellular carcinoma cells while also increasing cell death in those cells. The Rab protein family includes a sub-group known as Rab27, which is composed of RAB27A. In addition to that, it is believed that they play a role in controlling the progression of cancer.
It has been suggested that RAB27A plays a role in accelerating the course of breast cancer by encouraging cell invasion and metastasis. Additionally, it has been associated with gliomas and melanoma, suggesting that it may have a role as an oncoprotein in the development of human cancer.
According to the findings of the previous studies, the researchers found conflicting evidence on the involvement of the protein in mediating the advancement of colorectal cancer.
RAB27A was shown to have relatively high expression levels in the SW480 sample, but low expression levels in the RKO sample, according to the findings of a prior study carried out by the same authors. It was also shown that SW480 cells had a greater capacity for invasion, migration, and proliferation rate than RKO cells did. This led the researchers to believe that there was a positive correlation between RAB 27A and these other characteristics of SW480 cells.
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The researchers decided to use RKO and SW480 as cell models for RAB27A overexpression and knockdown, respectively.
In the experiment, the RAB27A shRNA plasmid was introduced into the SW480 cells, while the Flag-tag RAB 27A plasmid was introduced into the RKO cells. Researchers chose cell clones that stabilized the neo gene by using geneticin as a selection tool.
They discovered that the levels of expression of RAB274 mRNA and protein were much lower in the RAB 27A knockdown SW480 cells compared to the levels of expression in the control cells. In addition, an increase was seen in the levels when RKO cells that had been stably transfected were compared to the control cells.
In this work, the RAB27A knockdown SW480 cells had an optical density value that was noticeably lower compared to the negative control cells. This indicated that the knockdown limits cell growth. In addition, the optical density value in RKO cells was noticeably higher than that of the negative control cells, which suggested that RAB 27A ectopic expression encourages the proliferation of RKO cells.
In addition, the researchers used knockdown stable cell lines as well as ectopic expression cell lines to examine the impact that RAB27A had on the migration and invasion of cells.
They discovered that RAB27A knockdown cells were much lower than in the group that served as the negative control, which indicates that RAB 27A is necessary for the process of mediating the migration of colorectal cancer cells. It was demonstrated with RKO cells that RAB 27A is involved in the process of cell migration.
In addition, researchers discovered that RAB27A either prevented or aided overexpression in the cells depending on whether the cells were RKO or SW480. RKO cells invaded more easily than SW480 cells.
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Function of RAB27A ?
The RAB27A gene tells the body how to make a protein that moves proteins and other molecules inside cells in sac-like structures called vesicles. This process is called vesicle trafficking.