The Role Of RNA-binding Protein Involved In Stress Regulation In The Occurrence And Development Of Colorectal Cancer

Background: Colorectal cancer(CRC)has become the second most common disease related to tumor deaths and a global public health problem.In my country,the incidence of CRC has been increasing in recent years,and the number of new cases of CRC in 2020 has occupied the second place in the number of new cases of cancer.Although early-stage patients can have a good prognosis through surgery,chemotherapy,molecular targeting,and immunotherapy,the prognosis of patients with advanced colorectal cancer is still not ideal,and the mortality rate remains high.Its occurrence and development are regulated by complex multiple factors.Some unhealthy lifestyles,genetic factors,environmental factors and abnormal gene expression play an important role in the occurrence and development of CRC.The stress regulation mechanism involved in RNA binding protein is among them.An important component is still not comprehensive enough for its research.Further research on this process will help us understand the disease more comprehensively,and is of great significance for finding new CRC treatments and benefiting CRC patients.Methods: In order to prove that in colon cancer cells,different types of stress responses can also stimulate the formation of stress particles,accompanied by changes in RNA binding proteins,We constructed the corresponding cell stress model and observed whether the stress granules were produced.In order to understand the endogenous expression of RNA binding protein as an important component and regulatory factor of stress particles in colon cancer cells under stress,and the endogenous expression of RNA binding protein related to alternative polyadenylation regulation.We used Western blot to detect changes in the expression level of related RNA binding proteins under stress conditions after constructing a cell model of oxidative stress.Finally,in order to lay the foundation for further research on the influence of knockdown of related RNA binding proteins on stress regulation,the preliminary establishment of RNA binding protein knockout model was carried out by si RNA interference experiment.Results: When colon cancer cells are treated with Sodium Arsenite,heat shock,Dithiothreitol,and Tunicamycin,obvious TIA1 and TIAR fluorescence dot aggregations can be observed in the cytoplasm.After being treated with Sodium Arsenite and Tunicamycin for 2h,the aggregation of G3BP1 fluorescent spots can be observed.When processed by Sodium Arsenite,the PABPC1 fluorescent dots can be gathered.It is worth noting that in the absence of stimulating factors,exogenous overexpression of TIAR also found a small amount of TIAR fluorescent spot aggregation at 2h.After Sodium Arsenite treated colon cancer cells,the expression of endogenous TIA1increased;when treated for 0.5h,the endogenous expression of TIAR increased significantly,but after 2h,the expression of TIAR decreased slightly;after 0.5h,the expression of TIAR was slightly There is a decrease,and it only increases after 2 hours of treatment;the endogenous expression of CPEB1,CPEB3,and CPEB4 all increase to varying degrees TIA1 was successfully knocked down in the knockout model,and the TIAR knockout model was not successfully constructed.In the AUF1 knockout model,the expression of the three subtypes of AUF1 p45,AUF1 p42,and AUF1 p40 decreased,especially AUF1 p45 and AUF1 p40.The expression of AUF1 p37 subtype increased.Conclusions: 1.Exogenous overexpression of TIA1,TIAR,G3BP1,PABPC1 can induce colon cancer cells to produce stress granules under stress.2.In the absence of stress factors,the only overexpression of TIAR is sufficient for spontaneously inducing the assembly of stress particles.3.Under oxidative stress conditions,endogenous TIA1,TIAR,and G3BP1 will be highly expressed in colon cancer cells.4.The alternative polyadenylation process involving RNA-binding proteins is involved in the regulation of colorectal cancer stress.