ABSTRACT
Objective To detect the expression and function of ZNRF3 in different kinds of thyroid cancer tissues and cells.Methods Immunohistochemistry was used to detect the expressions of ZNRF3 protein in 35 cases of papillary thyroid carcinoma and 10 cases of poorly differentiated thyroid carcinoma.The expressions of ZNRF3 gene in TPC-1 and 8505C were detected by RT-PCR,and the cell lines were derived from papillary thyroid carcinoma and poorly differentiated thyroid carcinoma respectively.After silenced ZNRF3 gene expression with lentivirus,the proliferation ability of TPC-1 cells were detected with CCK-8,the invasion and metastasis ability of TPC-1 cells were detected with Transwell.Results According to results of immunohistochemistry and RT-PCR,the expressions of ZNRF3 in papillary thyroid carcinoma cells and tissues were higher than those in poorly differentiated thyroid carcinoma cells and tissues,the differences were statistically significant (4.83±0.44 vs.3.13 ±0.59,t =2.20,P ＜0.05;1.01±0.06 vs.0.21±0.04,t =11.80,P＜0.01).After ZNRF3 geng silencing,according to the results of CCK8,the proliferation ability of TPC-1 cells was significantly enhanced in 72 h,the difference was statistically significant (0.96 ± 0.10 vs.0.64 ± 0.05,t =3.19,P ＜ 0.05);and according to the results of Transwell,the TPC-1 cell's invasion (0.12 ± 0.01 vs.0.09 ±0.00,t =5.48,P＜0.01) and migration (0.22 ±-0.01 vs.0.17 ±0.01,t =4.58,P ＜0.05) also increased,the differences were statistically significant.Conclusion The expression of ZNRF3 in papillary thyroid carcinoma is higher than that in poorly differentiated thyroid cancer.ZNRF3 is tumor suppressor gene in the thyroid tumors.
ABSTRACT
Objective To investigate the effects of silencing of the human cerebral 21 .5 kDa myelin basic protein (MBP) gene on the proliferation and apoptosis of the glioma U251 cells .Methods The 21 .5 kDa MBP sequence‐specific short hair‐pin RNA (shR‐NA) recombinant plasmids pGenesil‐1‐MBP‐3 were transfected into the human glioma cell line(U251) ,the cells of U251 was used as MBP silencing group ,the cells transfected with negative control plasmids used as negative control group ,and the cells transfected with liposomes used as blank control group .Real‐time PCR and Westernblot were used to detect the expression levels of the 21 .5 kDa MBP mRNA and protein in each group ,and the cell proliferation curve was measured by CCK‐8 assay ,the apoptosis rate was a‐nalysised by Flow cytometry .Results Both the mRNA and the protein expression levels of the 21 .5 kDa MBP of MBP silencing group were significantly lower than those in the control groups (P<0 .05);the cellular proliferation activity of the MBP silencing group decreased significantly (P<0 .05)while the cellular apoptotic rate increased significantly (P<0 .05) .Conclusion Silencing of the human cerebral 21 .5 kDa MBP gene may playa dual role in the inhibition of proliferation and the promotion of apoptosis of the glioma U251 cells .
ABSTRACT
Objective: To investigate the inhibitory effect of short hairpin RNA (shRNA) -mediated silence of phospholipase C epsilon (PLC)̇gene expression on the proliferation of renal carcinoma 786-0 cells and its action mechanism. Methods: Liposome was employed to mediate the transfection of both the recombinant plasmid (pGenesil-PLC)̇and the control plasmid (pGenesil-NP) into the 786-0 cells. RT-PCR was used to detect the PLĊ mRNA expression after being transfected for 48 h. MIT assay was conducted to detect the proliferation inhibitory rate at 24, 48, and 72 h after transfection, respectively. Flow cytometry (FCM) was performed to analyze cell cycle after 48-h transfection. RT-PCR and immunocytochemistry were used to analyze the expression levels of both p27 and Ki67. Results: The expression of PLĊmRNA was significantly inhibited by recombinant plasmid transfection with the inhibitory rate of 69.7%. The proliferation inhibitory rates were 21.2%, 31.6%, and 32.7% after being transfected for 24, 48, and 72 h, respectively. FCM analysis demonstrated that the distribution of cell cycle changed. The number of cells in the G0/G1 phase increased, and that in both the S phase and G2/M phase decreased. Cells were arrested at the G0/G1 phase, and the subdiploid "apoptotic peak" appeared at the same time. RT-PCR and immucytochemistry indicated that the expression of p27 was up-regulated and that of Ki67 was downregulated. Conclusion: The proliferation of the renal carcinoma 786-0 cell line is inhibited by interference of PLĊgene expression, and the underlying mechanism may partially be related with up-regulation of P27 and down-regulation of Ki67.