The Study On The Relatives Between Alternative Splicing In VEGFA, APP And NUMB Gene And Colorectal Cancer

Objective:The purpose of this study was to investigate the relatives betweenalternative splicing in VEGFA, APP and NUMB gene and colorectal cancer.Methods:(1) Real-time quantitative reverse transcriptase polymerase chainreaction (real-time qRT-PCR) analysis was performed to detect theexpressions of VEGFA, APP and NUMB mRNA in20fresh CRC tissues andmatched normal intestinal mucosa tissues,in order to investigate the relativesbetween them and development and lymph node metastasis of colorectalcancer.(2) Restriction Fragment Length Polymorphism (RFLP) analysis wasperformed to detect the expressions of alternative splicing variants of VEGFA,APP and NUMB mRNA to explore the relatives between them and colorectalcancer.(3)In view of the expression of VEGFA alternative splicing variantVEGFA165b, its role in the development of colorectal cancer need toinvestigate.Results:(1) qRT-PCR analysis revealed that the expressions of APP and NUMBmRNA in CRC tissues were significantly lower than that in normalintestinal mucosa tissues, while VEGFA mRNA in CRC tissues was significantly higher than that in normal intestinal mucosa tissues(P <0.05).The expressions of them were uncorrelated with depth of tumor infiltration,the presence of lymph node metastasis and TNM stages.(2) RFLP analysis revealed that the expression of VEGFA alternativesplicing variant VEGFA165a/b in CRC tissues was significantly higher thanthat in normal intestinal mucosa tissues (P <0.05), the expression of VEGFAalternative splicing variant VEGFA121a/b was not statistically significantbetween CRC tissues and normal intestinal mucosa tissues(P>0.05), theexpression of VEGFA alternative splicing variant VEGFA206a/b was in fewsamples; The expressions of APP alternative splicing variants APP695,APP751and APP770were not statistically significant between CRC tissuesand normal intestinal mucosa tissues(P>0.05); The expression of NUMBalternative splicing variant NUMB-PRRSin CRC tissues was significantlylower than that in normal intestinal mucosa tissues (P <0.05), the expressionof NUMB alternative splicing variant NUMB-PRRLin CRC tissues wassignificantly higher than that in normal intestinal mucosa tissues (P <0.05).(3) qRT-PCR analysis revealed that the expression of VEGFAl65b inCRC tissues were significantly lower than that in normal intestinal mucosatissues (P <0.05). The expression of VEGFA165b were uncorrelated withdepth of tumor infiltration, the presence of lymph node metastasis and TNMstages.Conclusion:(1) VEGFA mRNA might play a role in promoting the development ofCRC in integrity, and VEGFA165a/b might play an important role among allthe VEGFA alternative splicing variants.(2)APP mRNA might play a role in inhibiting the development of CRC in integrity, while APP alternative splicing variants were not statisticallysignificant between CRC tissues and normal intestinal mucosa tissues.(3) NUMB-PRRSmight play a role in inhibiting the development of CRC,while NUMB-PRRLmight play a role in promoting the development of CRC.NUMB-PRRSappears to be more functional than NUMB-PRRL,as a consequence,NUMB mRNA plays a role in inhibiting the development of CRC in integrity.(4) VEGFA165b might play a role in inhibiting development of CRC,while VEGFA165a might play a role in promoting the development of CRC.Alternative splicing of mRNA provide a new target for the diagnosis,prognosis and treatment of this malignancy.