Effects Of Survivin ShRNA-APC Double Gene Co-Expression Stable Cell Strains On PERK-ATF4-CHOP Apoptosis Pathway Expression In Endoplasmic Reticulum Stress Of Colon Carcinoma

Objective:To investigate the effect of Survivin gene-specific short hairpin ribonucleic acid-colon adenomatous polyposis gene(Survivin sh RNA-APC double gene)on the expression of endoplasmic reticulum stress PERK-AFT4-CHOP apoptosis pathway in subcutaneous transplanted tumor tissue of HT-29 colon cancer cells in nude mice.Methods:Thirty healthy nude mice with no specific pathogen level were selected,and they were divided into 6 mice/group by random number table method.There are five groups:Survivin sh RNA-APC double genome,Survivin sh RNA single genome,APC single genome,no-load group and blank group(negative control group).Survivin sh RNA stable expression cell strains,APC stable expression cell strains,no-load cell strains and HT-29 colon cancer cells were constructed and validated earlier in this study.They were successively injected into the right anterior axilla of each group of nude mice,and kept in the same growth environment to continue feeding and observe the tumorigenesis of nude mice in each group.Six weeks later,all the nude mice were sacrificed by spinal cord dissection,and the tumorigenesis tissues were stripped,paraffin embedded and fixed to make tissue sections.Immunohistochemical technique and image pro plus6.0 image analysis software were used to determine the protein expressions of PERK,e IF-2α,GRP78,ATF4,Caspase-12,CHOP and Mean of IOD in subcutaneous transplanted tumor tissues of nude mice in each experimental group.The degree of cell apoptosis of transplanted tumor in each group was observe by TUNEL experiment,and the apoptosis index of each group was compared.Results:Observed the growth of subcutaneous transplanted tumor in nude mice of each group,and compared the average tumor volume of transplanted tumor at each observation time node.The average tumor weight after stripping tumor tissue was compared,and the tumor weight inhibition rate was calculated.The protein expressions of PERK,EIF-2 α,GRP78,ATF4,CHOP,caspase-12 and the apoptosis index of colon cancer cells in each group were statistically analyzed.(1)Contrasted with the negative control group and no-load group,the mean of tumor volume and the mean of tumor weight in Survivin sh RNA-APC double genome,Survivin sh RNA single genome and APC single genome were significantly reduced(P<0.05).Compared with single genome,the mean of tumor volume and the mean of tumor weight decreased more obviously in double genome(P < 0.05).There were no significant differences in mean tumor volume(P=0.660)and mean tumor weight(P=0.151)between the negative control group and the no-load group.(2)The tumor weight suppression rate of double genome(62.688%)is higher than that of APC single genome(37.153%)and Survivin sh RNA single genome(40.632%)(P < 0.05).(3)Immunohistochemical quantitative analysis of PERK,e IF-2α,GRP78,ATF4,CHOP,Caspase-12 proteins in transplanted tumor tissues of each group: the proteins in Survivin sh RNA-APC double genome,APC single genome and Survivin sh RNA single genome all expressed in different degrees,and the Mean of IOD increased(the protein expression content increased),and the increase of Mean of IOD in double genome was more significant than that in single genome.Compared with negative control group,the mean of IOD in no-load group had no significant difference(P < 0.05).(4)Statistical analysis of the results of apoptosis of colorectal carcinoma cells in each group: the apoptosis index of colorectal carcinoma cells in double genome(56.327±3.208),APC single genome(32.333±2.513)and Survivin sh RNA single genome(34.245±2.774)increased significantly.Compared with single genome,the apoptosis index of colorectal carcinoma cells in double genome has increased significantly(P < 0.05).Contrasted with the negative control group(9.928±0.297),the no-load group(10.162±0.577)had no significant difference(P=0.857).Conclusion:The Survivin gene-specific short hairpin ribonucleic acid-colon adenomatous polyposis gene(Survivin sh RNA-APC double gene)can significantly induce apoptosis of HT-29 colon cancer cells,which may play a synergistic role in inducing apoptosis of colon cancer cells by up-regulating the expression of endoplasmic reticulum stress PERKATF4-CHOP signaling pathway related proteins in colon cancer cells,and the regulation ability of double genomes is stronger than that of single genomes.