Proliferation Of TPC-1 Cells Research Articles

MYB/LINC00092 regulatory axis promotes the progression of papillary thyroid carcinoma.

Thyroid carcinoma is the most frequent malignancy in different endocrine-related tumours. In this study, we demonstrated a long non-coding RNA LINC00092-associated molecular mechanism in promoting the progression of papillary thyroid carcinoma (PTC). The expression of LINC00092 was analysed in the The Cancer Genome Atlas Thyroid Cancer (TCGA-THCA) patient cohorts and further determined by q-PCR. (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) (MTT) assay, and wound healing assay confirmed the function of LINC00092 in migration and proliferation. Q-ChIP validated the transcriptional target. Luciferase reporter assay validated the miRNA-mRNA target. The analysis in patient cohorts and in PTC TPC-1 cells showed that the expression of LINC00092 was repressed in thyroid carcinoma. In addition, the expression of LINC00092 was negatively associated with the advanced thyroid TNM stages. LINC00092 repressed epithelial-mesenchymal transition (EMT), migration, and proliferation of TPC-1 cells. Interestingly, we identified that MYB, a well-studied tumour promoter, is a transcription factor of LINC00092, thereby the expression of LINC00092 was directly repressed by MYB. Furthermore, miR-4741 was also validated as a direct target of MYB and was induced by MYB. Notably, LINC00092 was repressed by miR-4741 through the direct 3'-untranslational region (3'-UTR) target. Therefore, MYB induced EMT of TPC-1 cells by repressing LINC00092 directly or indirectly via miR-4741. Our study validated that LINC00092 is a tumour suppressor lncRNA in PTC. MYB directly or indirectly represses LINC00092, which contributes to the PTC progression. MYB, LINC00092, and miR-4741 form a coherent feed forward loop. The axis of MYB-LINC00092 promotes progression of PTC.

Circular RNA eukaryotic translation initiation factor 6 facilitates TPC-1 cell proliferation and invasion through the microRNA-138-5p/lipase H axis.

Both circular RNA eukaryotic translation initiation factor 6 (circEIF6) and microRNA (miR)-138-5p participate in thyroid cancer (TC) progression. Nevertheless, the relationship between them remains under-explored. Hence, this research ascertained the mechanism of circEIF6 in TC via miR-138-5p. After TC tissues and cells were harvested, circEIF6, miR-138-5p, and lipase H (LIPH) levels were assessed. The binding relationships among circEIF6, miR-138-5p, and LIPH were analyzed. The impacts of circEIF6, miR-138-5p, and LIPH on the invasive and proliferative abilities of TPC-1 cells were examined by Transwell and EdU assays. Tumor xenograft in nude mice was established for in vivo validation of the impact of circEIF6. CircEIF6 expression was high in TC cells and tissues. Additionally, miR-138-5p was poor and LIPH level was high in TC tissues. Mechanistically, circEIF6 competitively bound to miR-138-5p to elevate LIPH via a competitive endogenous RNA mechanism. Silencing of circEIF6 reduced TPC-1 cell proliferative and invasive properties, which was annulled by further inhibiting miR-138-5p or overexpressing LIPH. Likewise, circEIF6 silencing repressed the growth of transplanted tumors, augmented miR-138-5p expression, and diminished LIPH expression in nude mice. Conclusively, circEIF6 silencing reduced LIPH level by competitive binding to miR-138-5p, thus subduing the proliferation and invasion of TPC-1 cells.

Effects of Ursolic Acid in Extracts of Prunella vulgaris on the Proliferation of Thyroid Papillary Carcinoma Cells Under the p53MAPK Signaling Pathway

The aim of this research was to demonstrate the impact of ursolic acid (UA) in Prunella vulgaris extracts on the proliferation of papillary thyroid carcinoma (PTC) cells through the p53MAPK signaling. Effects of Prunella vulgaris extracts on TPC-1 cell proliferation were analyzed by intervening with various concentrations of UA, including negative control (NC) group, solvent control (SC) group, 3 μM UA group, 6 μM UA group, 12 μM UA group, and 15 μM UA group. Flow cytometry was adopted to evaluate apoptosis in TPC-1 cells, while real-time fluorescent quantitative (RT-q) PCR was implemented to assess expression (EP) of Bax and Bcl-2 in TPC-1 cells following UA intervention. RT-qPCR and Western blotting were employed to examine the differential EP levels of cell apoptosis, Bax, and Bcl-2 proteins. RT-qPCR was utilized to investigate the influence of UA on EP of various genes in MAPK pathway. The ethyl acetate extract exhibited the most notable inhibitory effect on TPC-1 cells. The content of UA in Prunella vulgaris increased gradually with the extension of ultrasonic time. The growth curve of TPC-1 cells demonstrated an initial increase followed by a decrease with increasing time. As the concentration increased, cell proportion in S phase increased, while the proportions in the GO-G1 and G2-M phases decreased, indicating that UA concentration-dependently arrested cells in the S phase. The level of Bax mRNA exhibited an increasing trend with increasing concentration, and the 12 μM UA and 15 μM UA groups demonstrated remarkable differences versus NC group (P <0.01). Bcl-2 protein demonstrated a decreasing trend with increasing concentration, and the 6 μM UA, 12 μM UA, and 15 μM UA groups exhibited considerable differences relative to NC group (P < 0.05). Additionally, pro-apoptotic protein Bax increased, while that of anti-apoptotic protein Bcl-2 decreased. UA treatment upregulated EP of the p53 gene in the MAPK pathway. Genes such as ERK, MEK, TSHR, Ras, p53, BRAF, PAK4, and PAKCa were downregulated. In summary, UA can upregulate EP of the p53 gene in the MAPK pathway, greatly inhibit proliferation of TPC-1 cells in PTC, and promote apoptosis. These findings provide insights for therapy of thyroid cancer.

MicroRNA-130a-3p impedes the progression of papillary thyroid carcinoma through downregulation of KPNB1 by targeting PSME3.

Papillary thyroid carcinoma (PTC) is the main type of thyroid cancer (THCA). Despite the good prognosis, some PTC patients may deteriorate into more aggressive disease, leading to poor survival. Our study aimed to explore the role of microRNA (miR)-130a-3p in regulating PTC. After transfection with miR-130a-3p-mimic, OE-PSME3, or miR-130a-3p-mimic + OE-KPNB1 in PTC cells (TPC-1), CCK-8, Transwell, scratch, and flow cytometry experiments were performed to analyze TPC-1 cell proliferation, invasion, migration, and apoptosis. Western blotting was used to detect proliferation or invasion-related protein markers (PCNA, E-cadherin, and N-cadherin). The RNA22 database, dual-luciferase reporter assay, and RNA pull-down assay were applied for the prediction and verification of the binding site between miR-130a-3p and PSME3. Pan-cancer software identified a positive correlation between PSME3 and KPNB1 in THCA. Co-immunoprecipitation was utilized to verify the interaction of PSME3 with KPNB1. Nude mice were transplanted with TPC-1 cells overexpressing miR-130a-3p. The tumors were isolated for detection of the expression of miR-130a-3p, PSME3, KPNB1, Ki-67, and CD31. miR-130a-3p was lowly expressed in PTC cell lines. Upregulation of miR-130a-3p repressed the expression of PSME3 and KPNB1 and reduced the malignancy of TPC-1 cells in vitro, shown by inhibited cell proliferation, invasion, migration, and the expression of PCNA and N-cadherin. Also, overexpressed miR-130a-3p inhibited the growth of xenograft tumors in nude mice. miR-130a-3p bound to PSME3 which interacted with KPNB1. miR-130a-3p impedes the progression of PTC by downregulating PSME3/KPNB1.

MiR-576-5p Promotes the Proliferation of Papillary Thyroid Carcinoma through the MAPK4-AKT Pathway.

MicroRNA-576-5p (miR-576-5p) plays an important role in different human cancers. However, the biological function of miR-576-5p in papillary thyroid carcinoma (PTC) is still unclear. In this study, we explored the function and specific role of miR-576-5p in PTC. Expression levels of miR-576-5p in PTC patient tissues and cell lines were determined by reverse transcription-quantitative polymerase chain reaction (qRT‒PCR). Cell counting using cell counting kit-8 (CCK-8), wound healing, and Transwell assays were performed to evaluate the effect of miR-576-5p on the proliferation, migration, and invasion of TPC-1 cells. Expression levels of mitogen-activated protein kinase 4 (MAPK4) and phosphorylation levels of protein kinase B (AKT), extracellular regulated protein kinase (ERK), and P38 mitogen-activated protein kinase (P38) were detected by western blotting or immunohistochemistry (IHC). The expression level of miR-576-5p in PTC tissues and TPC-1 cells was significantly increased. In vitro, overexpression of miR-576-5p promoted the proliferation, migration, and invasion of TPC-1 cells. In addition, MAPK4 was highly expressed in PTC tissues, and miR-576-5p could upregulate the expression of MAPK4. Interestingly, MAPK4 knockdown reversed cell proliferation but not migration and invasion in TPC-1 cells after miR-576-5p was overexpressed. Moreover, overexpression of miR-576-5p induced activation of the AKT pathway in TPC-1 cells, and MAPK4 gene knockout reversed this AKT pathway activation. In this study, we found that miR-576-5p was significantly overexpressed in PTC tissues and TPC-1 cells. In addition, miR-576-5p promoted the proliferation of TPC-1 cells by enhancing expression of MAPK4 and activating the AKT pathway.

Effects of Nanosecond Pulsed Electric Fields in Cell Vitality, Apoptosis, and Proliferation of TPC-1 Cells.

Objective To evaluate the effects of nanosecond pulsed electric fields (nsPEFs) with different pulse durations in cell vitality, apoptosis, and proliferation of TPC-1 cells, optimize pulse parameters and expand the application range of nsPEFs. Methods The pulse duration of 0, 300 ns, 500 ns, and 900 ns is generated with nsPEF generator. CCK-8 was used to investigate the effect of nsPEFs on the viability of TPC-1 cells. Flow cytometry was used to evaluate the apoptosis of TPC-1 after pulse treatment. The effect of nsPEFs on the proliferation ability of TPC-1 cells was detected by 5-ethy-nyl-2′-deoxyuridine. The morphological changes of TPC-1 cells after pulse treatment were observed by transmission electron microscopy. Results NsPEFs with 900 ns pulse duration can significantly affect the viability of TPC-1 cells and inhibit the proliferation ability of TPC-1 cells. In addition, nsPEFs can also induce apoptosis of TPC-1 cells. Conclusion NsPEFs with longer pulse duration can significantly affect the biological behavior of TPC-1 cells, such as cell viability and proliferation ability, and can also induce cell apoptosis, thereby inhibiting cell growth.

Overexpression of centromere protein K (CENPK) gene in Differentiated Thyroid Carcinoma promote cell Proliferation and Migration

ABSTRACT Differentiated thyroid carcinoma (DTC) is one of the most common malignant tumors. Increasing evidence indicates that centromere protein K(CENPK) may play a key role in promoting carcinogenesis. The expression, biological functions, and clinical significance of CENPK in DTC are still unclear. The CENPK expression in the DTC specimen was confirmed using quantitative real-time PCR and Western blot. The expression of CENPK was silenced and promoted by lentivirus-mediated transfection with shRNA sequences or CENPK plasmid targeting CENPK in TPC1 and FTC-133 cells, respectively. Colony formation, Cell Counting Kit-8 (CCK-8), Transwell invasion, and scratch assays were performed to assess the malignant biological properties of FTC-133 and TPC1 cells. Tumorigenicity assay was performed using C57BL/6 mice to explore the influence of CENPK on the growth of TPC1. The present work suggested that the expression of CENPK remarkably increased in follicular thyroid cancer and papillary thyroid cancer tissue samples at the mRNA level. Immunohistochemical staining also showed consistent results at the protein level. In addition, CENPK mRNA expression level showed great value in diagnosis of DTC. Knockdown of CENPK significantly inhibited the invasion and migration of TPC1 and FTC-133 cells. In contrast, CENPK overexpression promoted invasion and migration of TPC1 and FTC-133 cells. Knockdown and overexpression of CENPK showed consistent effect on DTC tumor growth and expression of Ki-67 invivo. Our results indicated that CENPK was evidently upregulated in DTC. Knocking down CENPK suppressed TPC1 cell proliferation, invasion and migration. Targeting the CENPK may be anovel therapeutic method for DTC.

ETS like-1 protein ELK1-induced lncRNA LINC01638 accelerates the progression of papillary thyroid cancer by regulating Axin2 through Wnt/β-catenin signaling pathway

ABSTRACT Papillary thyroid carcinoma (PTC) characterized by distant metastasis is a major public health issue among women worldwide. LncRNA LINC01638 is reportedly a critical oncogene in the development of certain cancers. However, the biological function of LINC01638 in PTC is currently unclear. The goal of this study was to identify LINC01638 expression level and its role in PTC progression. The expression of LINC01638 was detected applying qRT-PCR. CCK-8 assay, colony formation assay, immunofluorescence staining and flow cytometric analysis were performed to assess cell proliferation and cell cycle. In addition, cell migration and invasion were examined via wound healing assay, transwell assay and western blot analysis. We found that LINC0163 was upregulated in PTC cells compared with normal thyroid gland epithelial cell line Nthy-ori3-1. ELK1 could act as a transcription factor of LINC01638 and induce LINC01638 expression. LINC01638 silencing inhibited cell proliferation, migration and invasion, and obstructed the progress of TPC-1 cell cycle. LINC0163 silencing activated Axin2 while suppressing the expressions of β-catenin, Cyclin-D1 and c-MYC. Rescue experiment utilizing the transfection of Axin2 overexpression plasmid weakened LINC01638 overexpression-enhanced TPC-1 cell proliferation, metastasis, cell cycle progress and Wnt/β-catenin pathway. These results indicate that LINC0163 regulates PTC progression via inhibition of Wnt/β-catenin and activation of Axin2, which may develop into a novel therapeutic strategy for PTC treatment.

Long non-coding RNA DLEUI promotes papillary thyroid carcinoma progression by sponging miR-421 and increasing ROCK1 expression.

We investigated the role of long non-coding RNA DLEU1 (deleted in lymphocytic leukemia 1) in the progression of papillary thyroid carcinoma (PTC). DLEU1 levels were higher in PTC cell lines (BHP5-16, TPC-1,8505C, and SW1736) and patient tissues (n=54) than in a human thyroid follicular epithelial cell line (Nthy-ori3-1) or adjacent normal thyroid tissues. High DLEU1 expression correlated positively with lymph node metastasis and advanced clinical stages in PTC patients. Bioinformatics, dual luciferase reporter, and RNA pulldown assays confirmed that DLEU1 directly binds to miR-421. Moreover, bioinformatics and dual luciferase reporter assays showed that miR-421 directly binds to the 3’untranslated region of the rho-related coiled-coil kinase 1 (ROCK1) in TPC-1 cells. PTC patient tissues and cell lines showed high ROCK1 mRNA and protein levels as well as low miR-421 levels. CCK-8, flow cytometry, wound healing, and Transwell invasion assays demonstrated that DLEU1 silencing decreases TPC-1 cell proliferation, survival and progression, but they can be rescued by miR-421 knockdown or ROCK1 overexpression. DLEU1 knockdown in TPC-1 cells decreased in vivo xenograft tumor size and weight compared to controls in nude mice. These findings demonstrate that DLEU1 promotes PTC progression by sponging miR-421 and increasing ROCK1 expression.

Effect of actin related protein 2/3 complex subunit 2 gene silencing on the proliferation and apoptosis of papillary thyroid carcinoma TPC-1 cells

Objective To investigate the effect of actin related protein 2/3 complex subunit 2 (ARPC2) gene silencing on the biological characteristics of papillary thyroid carcinoma (PTC) TPC-1 cells through lentivirus-mediated RNA interference. Methods TPC-1 cells infected with nonsense short hairpin RNA (shRNA) sequence lentivirus (shCtrl) was used as the control group. TPC-1 cells infected with ARPC2 shRNA interference sequence lentivirus (shARPC2) was used as the experimental group, in which the expression of ARPC2 gene was specifically interfered. The effects of silencing the expression of ARPC2 gene on the proliferation of TPC-1 cells were detected by using methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, Western blot and colony formation test. Flow cytometry and Western blot were conducted to detect the effect of silencing ARPC2 gene on TPC-1 cells apoptosis and related proteins. Results shARPC2 could efficiently infect TPC-1 cells, and the expression efficiency of green fluorescent protein was over 85%. Compared with the control group, TPC-1 proliferation was inhibited in the experimental group. The ratio of S-phase cells in the experimental group was reduced compared with that in the control group [(14.79±0.21)% vs. (21.13±0.33)%, t = 27.77, P < 0.05]. The ratio of G1 and G2/M-phase cells in the experimental group was increased compared with that in the control group [G1 phase: (67.57±0.08)% vs. (62.06±0.36)%, t=25.56, P < 0.05; G2/M phase: (17.64±0.12)% vs. (16.91±0.17)%, t=6.154, P < 0.05]. Meanwhile, the expressions of cell cycle-related proteins CDK2, CyclinE and CyclinD were reduced in the experimental group. The number of clone formation in the experimental group was less than that in the control group, the difference was statistically significant [(10±2) vs. (161±6), t=9.011, P < 0.05]. In addition, the apoptotic ratio of cells in the experimental group was higher than that in the control group [(8.60±0.77)% vs. (4.08±0.40)%, t=9.011, P < 0.05]. Western blot showed that the expressions of anti-apoptotic factors p21 and bcl-2 were reduced in the experimental group, while the expression of pro-apoptotic factor bax was increased. Conclusion The interference with the expression of ARPC2 regulated by shRNA can inhibit the proliferation, and promote the apoptosis of PTC TPC-1 cells, indicating that ARPC2 may be a possible biological new target for the treatment of PTC. Key words: Thyroid neoplasms; RNA interference; Cell proliferation; Apoptosis

MicroRNA-1225-5p inhibits the development and progression of thyroid cancer via targeting sirtuin 3.

The objective of this study was to grab the expression levels and biological function of microRNA-1225-5p in human thyroid cancer. The miR-1225-5p expression in thyroid cancer tissue and cell lines was detected, followed by detecting the effects of overexpression of miR-1225-5p on the proliferation, apoptosis, migration and invasion of thyroid cancer cells. Moreover, the target relationship between miR-1225-5p and sirtuin 3 (SIRT3) was investigated. Besides, the effect of miR-1225-5p on the activation of Wnt/β-catenin pathway was elucidated. The miR-1225-5p expression was downregulated in thyroid cancer tissues and cell lines. Overexpression of miR-1225-5p significantly inhibited TPC-1 cell proliferation, increased cell apoptosis, and suppressed migration and invasion. In addition, SIRT3 was verified as a functional target of miR-1225-5p. SIRT3 expression was overtly increased in thyroid cancer tissues and cell lines. Overexpression miR-1225-5p and SIRT3 at the same time could significantly reverse the effects of miR-1225-5p overexpression alone on the malignant phenotypes of thyroid cancer cells. Furthermore, overexpression of miR-1225-5p significantly inhibited the activation of the Wnt/β-catenin pathway, which was remarkably reversed after overexpression of SIRT3. Our data reveal that downregulation of miR-1225-5p may promote tumor cell proliferation and metastasis in thyroid cancer via a direct targeting of SIRT3. Activation of the Wnt/β-catenin pathway may be a key mechanism to mediate the role of miR-1225-5p/SIRT3 axis in thyroid cancer. miR-1225-5p may serve as a potential anti-cancer target for thyroid cancer treatment.

FOXE1 supports the tumor promotion of Gli2 on papillary thyroid carcinoma by the Wnt/β-catenin pathway.

Papillary thyroid carcinoma (PTC) is a common malignancy in thyroid tissue. However, the molecular mechanism of PTC tumor progression remains unknown. The hedgehog (Hh) pathway is thought to play a key role during PTC development. Here we investigate the effects of glioma-associated oncogene protein-2 (Gli2), an important transcription factor of the Hh-signaling pathway, on PTC. Gli2 and forkhead box E1 (FOXE1) protein levels were upregulated in tissues of PTC patients and PTC cell lines. Using the PTC cell line TPC-1, we show that Gli2 small interfering RNA (siRNA) reduces cell proliferation, migration, and invasion; whereas overexpression of FOXE1 produces the opposite effects. Moreover, Gli2 siRNA inhibited the expression of genes implicated in the Wnt/β-catenin pathway and that FOXE1 overexpression produces the opposite effects. Thus, it was indicated that Gli2 promoted the proliferation, migration, and invasion of TPC-1 cells by activating Wnt/β-catenin and FOXE1 is involved in this process. Xenograft models of PTC were also constructed, the results showed that Gli2 siRNA reduced the rate of tumor growth, FOXE1 levels, and the expression of the Wnt/β-catenin pathway but FOXE1 overexpression reversed that effects. In conclusion, this study demonstrates that Gli2 promotes the growth of PTC tumors and TPC-1 cell proliferation, migration, and invasion by activating the Wnt/β-catenin pathway via FOXE1.

Mechanism of microRNA-1243 on proliferation and migration of human papillary thyroid carcinoma TPC-1 cells

Objective To observe the biological effects of microRNA (miRNA, miR)-1243 overexpression on proliferation and migration of human papillary thyroid carcinoma cell line TPC-1. Methods The expression of miR-1243 in 2×105 TPC-1 cells was detected by real-time quantitative polymerase chain reaction (RT-qPCR). The capacities of migration and the migration in TPC-1 cells were tested by water soluble methacon compounds (MTS) or Transwell assays, respectively. The candidate targets for miR-1243 were investigated using predication algorithm provided by miRanda and RNAhybrid. Luciferase activity was measured by dual luciferase reporter assay. Results The expression levels of miR-1243 in blank control and negative control groups were 0.97±0.31 and 1.208±0.40 respectively, significantly lower than 932.84±52.34 in miR-1243 mimic transfected group (P=0.000). The relative survival rates of the blank control group and the negative control group were 4.82±0.10 and 4.91±0.09, respectively, which were significantly higher than that of 4.41±0.06 in miR-1243 mimic transfected group (P=0.032). The proliferation ability of TPC-1 cells transfected with miR-1243 was significantly inhibited. The number of migratory cells in the miR-1243 mimic group was 15.5±6.5, which was significantly less than that in the blank control group (33.5±9.5) and the negative control group (31.75±10.75) (P=0.016). The bioinformatics analysis revealed that the AKT serine/threonine kinase 1 (AKT1) mRNA may be the target of miR-1243. The results of dual luciferase assay showed that WT 3’ untranslated region (UTR) of AKT1 cotransfected with miR-1243 mimics significantly decreased luciferase activity (45.92±10.91) as compared with empty vector group (98.97±9.96), mutant group (85.02±11.04) and negative control mimic group (97.38±10.29) (P=0.005). Conclusion Overexpression of miR-1243 can inhibit the proliferation and migration of TPC-1 cells. The miR-1243 may be involved in the regulation of thyroid papillary carcinoma by direct targeting of AKT1. Key words: Thyroid papillary carcinoma; MicroRNA-124; Cell proliferation; Cell migration; AKT serine / threonine kinase 1

Growth Inhibitory Effect of Mangiferin on Thyroid Cancer Cell Line TPC1

Thyroid cancer is the most frequently occurring cancer in human. Therefore, there is a need for better understanding and improvement in the treatment for thyroid tumorigenesis. Many studies show that natural dietary components like Mangiferin (present in mangoes) have anti-carcinogenic ability and reduce the incidence and development of cancers. The present study was focused on analysing the effect of Mangiferin on human thyroid cancer cell line (TPC1) and to assess its therapeutic significance against aggressive thyroid cancers. The cell viability of TPC1 cells was evaluated by MTT assay. The DAPI and AO/Er staining using fluorescence microscopy was used to evaluate the effects of the compound on apoptosis. The PCNA expression was also examined. In the present study, Mangiferin has significantly inhibits TPC1 cell proliferation. DAPI nuclear staining revealed apoptotic changes occurred in nuclei of cells treated with mangiferin (4 μM). AO/EtBr staining of TCP-1 cells showed that mangiferin treatment (2 and 4 μM) induced cell death. Mangiferin induces apoptosis through the induction of cas-3 and reduced Bcl-2 expression. It was concluded that mangiferin triggered the apoptotic pathways. It also decreased the viability of TPC-1 cells by suppressing the PCNA.

Matrine inhibits TPC-1 human thyroid cancer cells via the miR-21/PTEN/Akt pathway.

Papillary thyroid cancer (PTC) is the primary type of thyroid cancer and the most widespread endocrine malignancy. Matrine is a traditional Chinese medicine and has been demonstrated as a promising alternative drug for the treatment of TPC-1 human PTC. In the present study, the therapeutic effects and the underlying molecular mechanisms of matrine on TPC-1 cells were investigated. Treatment with matrine at the concentrations of 1, 2, 5, 10 and 20 mg/ml inhibited TPC-1 cell proliferation by up to 95.8% (for 20 mg/ml matrine). Flow cytometry indicated that treatment with 10 mg/ml matrine induced up to 61.8% apoptosis of the TPC-1 cells and the cell cycle was arrested at the G0/G1 phase following treatment with matrine (2, 5 and 10 mg/ml) for 48 h. Quantitative polymerase chain reaction indicated that the expression of microRNA (miR)-21 was downregulated and phosphatase and tensin homolog (PTEN) mRNA levels increased up to 1.66-fold following treatment with matrine, and RAC-α serine/threonine-protein kinase (Akt) mRNA levels were downregulated 0.34-fold following treatment with 5 mg/ml matrine, compared with the normal control group. Western blot analysis indicated that matrine at 2 and 5 mg/ml increased levels of the miR-21 target PTEN and decreased the levels of phosphorylated (p)Akt. Furthermore, miR-21 mimic transfection decreased the expression levels of PTEN and increased the levels of pAkt. These results suggested that the miR-21/PTEN/Akt pathway may be one of the mechanisms by which matrine induces apoptosis and cell cycle arrest in TPC-1 thyroid cancer cells. Matrine is an alternative potential drug for the treatment of thyroid cancer.

MicroRNA-625-3p promotes the proliferation, migration and invasion of thyroid cancer cells by up-regulating astrocyte elevated gene 1

BackgroundThyroid cancer is the most common malignancy in human endocrine system. This study aimed to investigate the effects of microRNA-625-3p (miR-625-3p) on thyroid cancer cell proliferation, migration, invasion and apoptosis, as well as underlying potential mechanism. MethodsThe relative expressions of miR-625-3p in tumor tissues and adjacent normal tissues of 20 patients with papillary thyroid cancer (PTC) were assessed using qRT-PCR. Cell transfection was used to up-regulate or down-regulate the expressions of miR-625-3p in thyroid cancer SW579 and TPC-1 cells. Effects of miR-625-3p overexpression or suppression on SW579 and TPC-1 cell viability, migration, invasion and apoptosis were detected respectively. The regulatory effect of miR-625-3p on astrocyte elevated gene 1 (AEG-1) expression was also analyzed. Then, the roles of AEG-1 in SW579 and TPC-1 cell proliferation, migration, invasion and apoptosis, as well as Wnt/β-catenin and c-Jun N-terminal kinase (JNK) pathways activation, were evaluated. ResultsmiR-625-3p had high expressions in tumor tissues, compared to adjacent normal tissues. Overexpression of miR-625-3p significantly promoted SW579 and TPC-1 cell proliferation, migration and invasion but had no influence on cell apoptosis. Knockdown of miR-625-3p had opposite effects, but induced cell apoptosis. AEG-1 was up-regulated by miR-625-3p overexpression and participated in the effects of miR-625-3p on SW-579 and TPC-1 cells. In addition, overexpression of AEG-1 induced the activation of Wnt/β-catenin and JNK pathways in SW579 and TPC-1 cells. ConclusionmiR-625-3p promoted proliferation, migration and invasion of thyroid cancer cells by enhancing the expression of AEG-1 and activating downstream Wnt/β-catenin and JNK pathways.

The expression of microRNA-150 in papillary thyroid carcinoma and its effect on proliferation and apoptosis

Objective To study the expression of miR-150 in papillary thyroid carcinoma (PTC) and the effects of miR-150 on the biological behavior of TPC-1. Methods The expression levels of miR-150 in PTC tissues and adjacent non-tumorous tissues were detected by real-time fluorescence quantitative PCR. Liposome 2000 was used to stably transfect miR-150 mimics and inhibitor respectively into TPC-1 cells. The cell counting kit-8 (CCK-8) method and flow cytometry were applied to analyze the effects of miR-150 on proliferation and apoptosis of TPC-1 cells. Results The expression level of miR-150 in PTC tissues (0.573 8±0.050 9) was significantly lower than that in the adjacent non-tumorous tissues (1.768 0±0.198 0) and the difference was statistically significant (P=0.000), and the expression level of miR-150 decreased significantly with the increase of staging of tumor (P=0.005). The CCK-8 results showed that proliferation ability of TPC-1 cells after transfection with miR-150 was significantly lower than that in the negative control group or the scrambled group (P=0.001). There was no statistically significant difference between the negative control group and the scrambled group (P=0.070). The apoptosis rate in miR-150 group [(14.79±1.85)%] was significantly higher than that in the NC group [(4.90±0.51)%] and the scrambled group [(5.79±1.00)%] (P=0.013), and there was no statistical difference between the the negative control group and the scrambled group (P=0.472). Conclusion miR-150 can inhibit the proliferation and apoptosis abilities of PTC cells and amy play an important role in the development of PTC. Key words: Papillary thyroid carcinoma; MicroRNA-150; Proliferation; Apoptosis

TFF3 Contributes to Epithelial-Mesenchymal Transition (EMT) in Papillary Thyroid Carcinoma Cells via the MAPK/ERK Signaling Pathway.

: Trefoil factor 3 (TFF3) was found to be overexpressed in many types of tumours. Evidence has shown that TFF3 plays an important role in tumour proliferation, migration and invasion metastasis. However, the impact of TFF3 on patients' clinicopathological characteristics and underlying mechanisms remain unknown in papillary thyroid carcinoma (PTC). In this study, the expression of TFF3 and the epithelial-mesenchymal transition (EMT) transcriptional factor Snail in PTC and para-carcinoma specimens was evaluated by immunohistochemistry (IHC) and Western blot, and the possible associations with lymph node (LN) metastasis and other clinicopathological parameters were analysed. In vitro, the effect of TFF3 on the malignant behaviour of TPC-1 cells was evaluated by cell proliferation assays, cell adhesion assays, colony formation assays, wound-healing assays and transwell chamber invasion assays. EMT markers and regulatory molecules were detected by quantitative RT-PCR (qRT-PCR) and Western blot analysis in the TFF3-knockdown groups and shRNA control group. The results showed that TFF3 was upregulated in PTC tissue and was associated with lymph node metastasis (P=0.0001), pathological grade (P=0.0002) and Snail expression (P=0.0001). The knockdown of TFF3 markedly inhibited the abilities of TPC-1 cell proliferation, adhesion, colony formation, migration and invasion. Mechanically, the results demonstrated that TFF3 might activate the MAPK/ERK signalling pathways, affect the expression of the transcription factors snail and slug in addition to affecting EMT associated markers E-cadherin and N-cadherin, and accelerate the progression of EMT in TPC-1 cells. These findings indicate that TFF3 might promote the metastatic potential of PTC by promoting the EMT process through cascades of MAPK/ERK pathways.

Reactivity of thyroid papillary carcinoma cells to thyroid stimulating hormone-dominated endocrine therapy.

This study investigated the effect of thyroid stimulating hormone (TSH) on the proliferation of papillary thyroid carcinoma (PTC) cells and the therapeutic effect of levothyroxine sodium (TH). PTC cells (TPC-1) were cultured using 0.1, 1.0 and 10 U/l TSH and 10−2, 10−4 and 10−6 mol/l TH. After the appropriate concentration was screened, TPC-1 cells were further divided into control group, TSH group, TH group and TSH+TH group. The cell proliferation was detected via methyl thiazolyl tetrazolium (MTT) method, TPC-1 cell cycle was detected via flow cytometer, and the mRNA and protein expression of cyclin D1 were detected via real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Compared with control group, TSH significantly promoted the proliferation of TPC-1 cells (P<0.05 or P<0.01), obviously promoted the transition of TPC-1 cells from G1 phase to S phase (P<0.01) and remarkably increased the mRNA and protein expression of cyclin D1 (P<0.01); but TH had a significant inhibitory effect on these results of TSH (P<0.05 or P<0.01). TSH can promote the proliferation of PTC cells, and the appropriate complement of TH can inhibit its proliferation.

Potential Antitumor Effect of Harmine in the Treatment of Thyroid Cancer.

Thyroid cancer is one of the most common types of cancer in endocrine system. In latest studies, harmine has been proved to inhibit the growth of several cancers in vitro and in vivo. In the current study, we evaluated the in vitro and in vivo anticancer efficiency of harmine against thyroid cancer cell line TPC-1. The in vitro cytotoxicity of harmine evaluated by XTT assay indicated that harmine suppressed the proliferation of TPC-1 cells in a dose- and time-dependent manner. Moreover, harmine dose-dependently induced apoptosis of TPC-1 cells through regulating the ratio of Bcl-2/Bax. The colony forming ability of TPC-1 cells was also time-dependently inhibited by harmine. The inhibitory effects of harmine on migration and invasion of TPC-1 cells were studied by wound scratching and Transwell assays. In vivo evaluation showed that harmine effectively inhibited the growth of thyroid cancer in a dose-dependent manner in nude mice. Therefore, harmine might be a promising herbal medicine in the therapy of thyroid cancer and further efforts are needed to explore this therapeutic strategy.