Effect of HE4 and PAX8 gene knockdown on malignant biological behaviors of epithelial ovarian cancer cells treated with TC regimen / 中国肿瘤生物治疗杂志

@#Objective: To investigate the effect of human epididymal protein 4 (HE4) and paired box gene 8 (PAX8) gene knockdown on proliferation, migration, invasion and apoptosis of human epithelial ovarian cancer OVCAR3 cells treated with TC regimen (paclitaxel+carboplatin). Methods: Sequences of single-target siRNA (HE4-siRNA or PAX8-siRNA) and double-target siRNA (HE4+PAX8siRNA) as well as negative siRNAwere respectively designed and synthesized, and then linked with plasmid vector pGCsi-H1 to obtain the recombinant plasmids. The obtained recombinant plasmids were then transfected into human epithelial ovarian cancer OVCAR3 cells, namely HE4-siRNA group, PAX8-siRNA group, HE4+PAX8-siRNA group and siRNA-NC group, respectively. The blank control group was also set up (without any treatment). The cells in above five groups were treated with TC regimen (paclitaxel 3.13 g/ml+carboplatin 2.82 µg/ml), and the changes in proliferation, migration, invasion and apoptosis of the cells were detected by MTT, wound-healing assay, Transwell chamber assay, and flow cytometry, respectively. Results: After knocking down the HE4 and PAX8 genes, compared with siRNA-NC group and blank control group, the proliferation, migration and invasion abilities of OVCAR3 cells in HE4-siRNA group, PAX8-siRNA group and HE4+PAX8-siRNA group significantly decreased (all P<0.01), and the apoptosis rate significantly increased (P<0.01), especially in HE4+PAX8-siRNA group. Conclusion: Knockout of either HE4 or PAX8 can enhance the effect of TC regimen on inhibiting proliferation, migration and invasion as well as promoting apoptosis of epithelial ovarian cancer cells, and the effect of simultaneous down-regulation of HE4 together with PAX8 is better.

Up-regulation of Bcl2l14 gene in myocardium of Pax-8 gene knockout mouse / 中国病理生理杂志

AIM: To investigate the downstream genes of the transcriptional factor Pax-8 related to cardiopathy. METHODS: The total RNA derived from the heart of Pax-8 KO~(-/-) and Pax-8 KO~(+/-) mice was extracted. Mouse genome DNA microarray containing 31 802 mouse oligonucleotides probes was used to investigate the differential expression between the Pax-8 KO~(-/-) and the Pax-8 KO~(+/-) mice hearts. The candidate genes were confirmed by RT-PCR and real time RT-PCR assay. RESULTS: Microarray results showed that, compared to the Pax-8 KO~(+/-) mice, 25 genes were down-regulated and 17 were up-regulated in the Pax-8 KO~(-/-) mice, concerning metabolize enzymes, cell signal conducting and nuclear transcript factors and so on. Bcl2-like 14 (Bcl2l14) was proved to be up-regulated by RT-PCR. Real time RT-PCR results revealed that Bcl2l14 in the Pax-8 KO~(-/-) mice was 2.07 and 2.23 fold as much as that in the Pax-8 KO~(+/-) and the Pax-8KO~(+/+) mice (P<0.01). CONCLUSION: The Bcl2l14 gene is one of the downstream genes of Pax-8 and probably plays an important role in the mechanism of ventricular septum defect.