Study Of The SH3-domain GRB2-like2Expression In Pancreatic Carcinoma

Background and Objective：Pancreatic carcer is a common abdominal highly malignant tumor.Along withdeep research on human genetics,people realized that the activation of oncogene andtumor suppressor gene inactivation and some other tumor gene mutation can cause theoccurrence of the tumor.SH3GL2gene is a kind of potential of TSG,which is widelyexpressed in human body.However,it is mainly expressed the central nervoussystem,and show significant down-regulation between the other tissues.Recentresearch showed that the significant low expression of SH3GL2on the followingtissues:human brain glioma,laryngeal cancer,breast cancer,stomach cancer andovarian cancer, however, the expression of SH3GL2on pancreatic carcinoma has notbeen declared yet.In the present study,we searched the expression of SH3GL2in both mRNA andprotein levels on three pancreatic cancer cell lines with different ability of invasionand metastasis (BxPC-3,PANC-1and Mia PaCa-2) and one normal pancreatic cellline (H6C7). Further more,the protein expression of SH3GL2on the differentdifferentiation degrees of pancreatic cancer specimens of paraffin wax were detectedby the method of immunohistochemistry,and analyzed that the relationship betweenthe expression of SH3GL2and the clinical pathological features in pancreaticcancer.Initially analyzed the expression of SH3GL2in pancreatic cancer, and revealedthat the relationship between the expression of SH3GL2and the differentdifferentiation degrees of pancreatic cancer tissues with the clinical pathologicalfeatures in pancreatic cancer.In order to lay the foundation for further research of themechanism of SH3GL2in invasion and metastasis of pancreatic cancer.Methods：1. A rapid and reliable real-time quantitative reverse transcription polymerase chain reaction(real time QPCR) approach using LightCycler Technology was used todetect the express of SH3GL2on three pancreatic cancer cell lines with differentability of invasion and metastasis (BxPC-3,PANC-1and Mia PaCa-2) and one normalpancreatic cell line(H6C7) in mRNA levels.2. Western blot was used to detect the expression of SH3GL2on three pancreaticcancer cell lines with different ability of invasion and metastasis (BxPC-3,PANC-1and Mia PaCa-2) and one normal pancreatic cell line(H6C7) in protein levels.3. Immunohistochemistry method was used to detect the expression of SH3GL2on the different differentiation degrees of pancreatic cancer tissues and the normalpancreatic tissues.4. Finally,analysed the relationship between the expression of SH3GL2and theclinical pathological features in pancreatic cancer.Results：1. In real time QPCR examination,three pancreatic cancer cell lines withdifferent ability of invasion and metastasis (BxPC-3,PANC-1and Mia PaCa-2) andone normal pancreatic cell line(H6C7) express the mRNA of SH3GL2.Compared to_β-actin,use x±s as measured2-ΔΔC(t)value,the2-ΔΔC(t)of SH3GL2in BxPC-3was14.330±0.254;the2-ΔΔC(t)of SH3GL2in PANC-1was7.768±0.341;the2-ΔΔC(t)ofSH3GL2in Mia PaCa-2was1.008±0.007;the2-ΔΔC(t)of SH3GL2in H6C7was16.284±0.216;the average compared with single factor variance analysis;P<0.05forthe difference was statistically significant.the expression of SH3GL2in all kinds ofcells is:H6C7> BxPC-3> PANC-1> Mia PaCa-2.2. The expression of SH3GL2in protein were confirmed by western blot.As theresult of real time QPCR,the expression of SH3GL2in all kinds of cells is:H6C7>BxPC-3> PANC-1> Mia PaCa-2.3. In immunohistochemistry examination, the expression of SH3GL2on threepancreatic cancer tissues with different differentiation degrees and one normalpancreatic tissue in protein levels. With lower degree of differentiation of pancreaticcancer, SH3GL2down the more obvious. Main orientation of SH3GL2proteins in the cytoplasm, consistent with earlier studies.4. It shows significant difference of SH3GL2in pancreatic cancer tissuescompared with the normal pancreatic tissues.The expression of SH3GL2waspositively correlated with the histological differentiation and TNM stage (p<0.05),butnot with the age,sex,tumor size, lymph node metastasis(p>0.05).Conclusion：1. The expressions of SH3GL2mRNA and protein are downregulated in all threepancreatic cancer cell lines. And the lower differention the tumor is, the lowerexpression of the SH3GL2.2. The expression of SH3GL2is downregulated in pancreatic cancer tissues,andits expression have correlation between the tumor differentiation and TNM stage.