ABSTRACT
Objective:To investigate the effects of miR-497 on cytobiology behaviors of papillary thyroid carcinoma (PTC) by targetingly regulating the expression of Yes-associated protein 1 (YAP1) .Methods:Human TPC-1 cells were divided into control group, miR-497 group, si-YAP1 group and miR-497+si-YAP1 group. The liposome transfection was conducted by LipofectamineTM3000. The targeted relationship between miR-497 and YAP1 was validated by Luciferase Reporter Assay. The cell proliferation activity in each group was detected by MTT method. The apoptosis rates were analyzed by flow cytometry. The number of invasion cells was detected by Transwell. The cell migration rates were detected by scratch assay. The expression of Cyclin D1, matrix metalloproteinase 2 (MMP-2) , MMP-9, matrix metalloproteinase inhibitor-1 (TIMP-1) and activated caspase 3 (cleaved Caspase-3) was detected by Western blot. SPSS 22.0 was used to analyze data, and normally distributed measurement data were expressed as ( ± s) . One-way ANOVA was analyzed for the difference between multiple groups, and SNK-q were analyzed for the difference between two groups. Results:Compared with the control group, the expression of YAP1 mRNA and protein was decreased in miR-497 group and si-YAP1 group ( q=14.682, 14.597; 23.743, 23.571; P<0.05) , cell proliferation activity, number of invasion cells and migration rate were decreased ( q=4.724, 4.568, 3.841; 4.216, 3.952, 3.274; P<0.05) , apoptosis rate was increased ( q=3.783; 4.336; P<0.05) , expression of CyclinD1, MMP-2, MMP-9 and cleared Caspase-3 proteins was decreased ( q=5.823, 5.981, 6.036, 6.485; 5.934, 6.110, 6.573, 6.614; P<0.05) , and expression of TIMP-1 protein was increased ( q=6.071; 6.148; P<0.05) . Compared with si-YAP1 group, miR-497 level was increased in miR-497+si-YAP1 group ( q=14.726, P<0.05) , the expression of YAP1 mRNA and protein was decreased ( q=3.089, 3.126; P<0.05) , cell proliferation activity, number of invasion cells and migration rate were decreased ( q=2.654, 2.537, 2.246; P<0.05) , apoptosis rate was increased ( q=2.875, P<0.05) , expression of CyclinD1, MMP-2, MMP-9 and cleared Caspase-3 proteins was decreased ( q=4.371, 4.365, 4.383, 4.368; P<0.05) , and expression of TIMP-1 protein was increased ( q=4.275, P<0.05) . Conclusion:MiR-497 can negatively targetingly regulate the expression of YAP1, inhibit proliferation, invasion and migration of TPC-1 cells.
ABSTRACT
Objective To detect the expression levels of vascular endothelial growth factor D ( VEGF-D) , D2-40 and MMP-7 in papillary thyroid carcinoma ( PTC) and to explore their relationship with micro lymphat-ic density, distribution,and cervical lymph node metastasis .Methods SP immunohistochemistry was used to study the expression of VEGF-D, D2-40 and MMP-7 in 38 cases of PTC with cervical lymph node metastasis , 45 cases of PTC without cervical lymph node metastasis , and 22 cases of normal thyroid tissues .Micro lymphatic density ( MLD) was counted .Results The expression level of VEGF-D in PTC with cervical lymph node metasta-sis and normal thyroid tissues had significant difference (χ2 =13.074, P=0.000, P<0.0167).The expression level of D2-40 in PTC with and without cervical lymph node metastasis had significant difference ( P<0.05 ) . MMP-7 expression in PTC with and without cervical lymph node metastasis had statistical significance ( 91.67%vs 73.33%, P<0.05).MLD in PTC with and without cervical lymph node metastasis and in normal thyroid tis-sues had statistical difference (11.7 ±3.5 vs 8.9 ±3.1 vs 3.9 ±2.7, P<0.05).Conclusion VEGF-D, D2-40 and MMP-7 are highly expressed in PTC and they are possibly related with cervical lymph node metastasis of PTC.