Expression analysis of TRAF2­ and NCK­interacting protein kinase (TNIK) and phosphorylated TNIK in papillary thyroid carcinoma.

The aim of the present study was to evaluate the expression of TRAF2- and NCK-interacting kinase (TNIK) and the levels of the active form of TNIK, phosphorylated (p)-TNIK, in papillary thyroid carcinoma (PTC), and to identify and compare the levels of TNIK and p-TNIK among PTC, benign thyroid tumors and normal tissues. The levels of TNIK and p-TNIK were examined by reverse transcription-quantitative (RT-q)PCR and immunohistochemical analysis (IHC) in PTC, benign thyroid tumors and normal tissues, and their association with clinicopathological features was evaluated. First, analysis of the Gene Expression Profiling Interactive Analysis and The Cancer Genome Atlas datasets suggested that the mRNA expression of TNIK was markedly increased in PTC tissues compared with that in normal tissues. RT-qPCR analyses then indicated that the relative mRNA expression of TNIK in PTC tissues was 4.47±6.16, which was significantly higher than that in adjacent tissues 2.57±5.83. The IHC results suggested that the levels of TNIK and p-TNIK in PTC tissues were markedly elevated compared with those in benign thyroid tumors and normal tissues. The levels of p-TNIK in patients with PTC were significantly associated with extrathyroidal extension (χ2=4.199, P=0.040). Positive staining for TNIK was observed in 187 out of 202 (92.6%) cases in the cytoplasm, nucleus or cytomembrane of PTC cells. Among the 187 positive cases, cytoplasm expression was identified in 162 cases (86.6%), nuclear expression in 17 cases (9.1%) and cytomembrane expression in 8 cases (4.3%). Positive staining for p-TNIK was observed in 179 out of 202 (88.6%) cases in the nuclei, cytoplasm or cytomembrane of PTC cells. In the 179 p-TNIK-positive cases, localization in the nuclei plus cytoplasm was identified in 142 cases (79.3%), nuclear localization in 9 cases (5.0%), presence in the cytoplasm in 21 cases (11.7%) and cytomembrane localization in 7 cases (3.9%). Both TNIK and p-TNIK were upregulated in PTC tissues and p-TNIK was significantly associated with extrathyroidal extension. It may act as a crucial oncogene to participate in PTC carcinogenesis and progression.

Hsa_circ_0008234 facilitates proliferation of cutaneous squamous cell carcinoma through targeting miR-127-5p to regulate ADCY7.

Cutaneous squamous cell carcinoma (cSCC) is the second most common skin malignant tumor with 25-50% of 5-year survival. There exist urgent needs for the identification of novel biomarkers for the diagnostic and therapeutic strategies of cSCC. The differentially expressed circRNAs in cSCC tissues and non-lesional skin tissues were obtained through analyzing the circular RNAs (circRNAs) microarray dataset GSE74758. The expression pattern of the indicated circRNAs in cSCC tissues was confirmed by qRT-PCR. FISH analysis was used to detect the location of hsa_circ_0008234 in cells. RIP experiment was used to detect the interaction between hsa_circ_0008234 and miR-127-5p. CCK-8 analysis and colony formation assay were used to detect the proliferation of cSCC cells. qRT-PCR and western blot were adopted to detect the expression of ACDY7. Three differential expressed circRNAs were obtained from the microarray data (GSE74758), and hsa_circ_0008234 was confirmed to be highly expressed in cSCC tissues by qRT-PCR. Hsa_circ_0008234 was mainly located in cytoplasm and stable in cSCC cells. RIP experiment revealed that hsa_circ_0008234 directly interacts with miR-127-5p in cSCC cells. Hsa_circ_0008234 increased the cell viability and colony formation of cSCC cells through acting as the sponge of miR-127-5p. MiR-127-5p inhibited the expression of ADCY7 in cSCC cells through binding the 3'UTR of ADCY7. Hsa_circ_0008234 was positively associated with ADCY7 expression in cSCC tissues. Hsa_circ_0008234 facilitates the proliferation of cSCC through targeting miR-127-5p to regulate ADCY7 expression and has the potential to be a novel therapeutic target for cSCC.

Expression of transcription factor FOXP4 in laryngeal squamous cell carcinoma tissues and its effects on biological characteristics of laryngeal squamous cell carcinoma TU177 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective:To detect the expression of transcription factor FOXP4 (Forkhead box P4) in laryngeal squamous cell carcinoma (LSCC) tissues and cell lines, and to investigate its effects on the proliferation, migration, invasion, cell cycle, and apoptosis of LSCC TU177 cells in vitro as well as to explore its relationship with epithelial-mesenchymal transition (EMT) process. Methods: A total of 40 pairs of tumor tissues and adjacent tissues that resected from LSCC patients were collected from the biological specimen bank of the Forth Hospital of Hebei Medical University between 2013 and 2015. The expression of FOXP4 in LSCC tissues and corresponding adjacent tissues was detected by qPCR. qPCR and Western blotting were used to detect the FOXP4 expression level in human LSCC cell lines (AMC-HN-8, TU177, TU686, and TU212). Small interfering RNA (si-RNA) was used to knock down FOXP4 expression in TU177 cells. The effects of FOXP4 knockdown on the proliferation, migration, invasion, cell cycle and apoptosis of TU177 cells were measured by MTS assay, clone formation assay, Transwell chamber migration and invasion assay, and flow cytometry, respectively. The mRNA levels of EMT markers N-cadherin, β-catenin, Vimentin, Twist, Snail and zine finger E box binding homeobox 1 (ZEB1) after transfection of si-FOXP4 in TU177 cells were detected by qPCR. The changes of protein levels of N-cadherin, β-catenin, Vimentin and Twist after FOXP4 knockdown were measured by Western blotting. Results: The expression of FOXP4 in LSCC tissues was significantly higher than that in adjacent tissues (P<0.05), and it was related to the TNM stage of tumors and lymph node metastasis (all P<0.05). The expression of FOXP4 in LSCC cells was higher than that in the adjacent tissues (P<0.05 or P<0.01). The expression of FOXP4 in TU177 cells transfected with si-FOXP4 was significantly lower than that in the control group (P<0.01). Compared with the control group, knocking down FOXP4 could inhibit the proliferation, migration and invasion but promote the apoptosis of TU177 cells in vitro (all P<0.01), block the cell cycle at G0/G1 phase (P<0.01), and reduce cell replication in S phase (P<0.01); in addition, knocking down FOXP4 could reduce the mRNA levels of N-cadherin, β-catenin, Vimentin, Twist, Snail, ZEB1 (P<0.05 or P<0.01) and the protein levels of N-cadherin, β-catenin, Vimentin, Twist in TU177 cells. Conclusion: The high expression of FOXP4 may be related to the occurrence and development of LSCC. FOXP4 knockdown can inhibit the proliferation, migration and invasion of laryngeal cancer cells in vitro, block cell cycle at G0/G1 phase, promote apoptosis, and may participate in the EMT process.

[Role of carbon nanoparticles in patients with thyroid carcinoma undergoing total thyroidectomy plus bilateral central neck dissection].

OBJECTIVE: To evaluate the role of carbon nanoparticles for dissecting lymph nodes and preserving parathyroid glands in patients with thyroid carcinoma undergoing total thyroidectomy plus bilateral central neck dissection. METHODS: A total of 86 patients with thyroid carcinoma undergoing primarily total thyroidectomy plus bilateral central neck dissection were randomly divided into trial and control groups. Carbon nanoparticles were injected into thyroid gland of trial group. Total lymph node, metastasis lymph node, black stained lymph node and black stained metastasis lymph node of trial group were counted in central compartment dissection specimens. And total lymph node and metastasis lymph node of control group were counted in central compartment dissection specimens. Parathyroid glands in central neck dissection specimens were counted in two groups. For two groups, serum total calcium and parathyroid hormone were measured pre-operation and 1 day, 3 days, 5 days, 1 month, 3 months and 6 months post-operation. RESULTS: In trial group, the average counts of (10.19 ± 4.27) lymph nodes and (8.44 ± 4.31) black stained lymph nodes were more than those in control group at (6.26 ± 2.98) lymph nodes (all P < 0.01). Parathyroid gland was found in trial group (n = 7) and control group (n = 11). And the difference had no statistical significance (χ(2) = 1.124, P = 0.289). The preoperative serum levels of calcium and parathyroid hormone decreased within 6 months postoperatively in both groups (all P < 0.01). Without extracapsular invasion, serum levels of calcium and parathyroid hormone were higher in trial group than those in control group at 5 days, 1 month and 3 months post-operation (all P < 0.05). In an event of lymph node metastasis, serum levels of calcium (t = 3.446, P = 0.001) and parathyroid hormone (t = 2.441, P = 0.017) in trial group were higher than those in control group at 1 month post-operation. When there was extracapsular invasion or no lymph node metastasis, the serum levels of calcium and parathyroid hormone had no inter-group statistical differences within 6 months post-operation (all P > 0.05). When tumor size was less than or equal to 4 cm, the level of parathyroid hormone was higher in trial group than that in control group at 1 month post-operation (t = 2.703, P = 0.009). But no inter-group statistical differences existed within 6 months post-operation (all P > 0.05) when tumor size surpassed 4 cm. Regardless of tumor size, the serum levels of calcium in trial group were higher than those in control group at 1 month post-operation (all P < 0.01). CONCLUSIONS: For thyroid carcinoma patients, total thyroidectomy plus bilateral central neck dissection increases the incidence of hypoparathyroidism. Lymph nodes of central compartment may be tagged by carbon nanoparticles so as to boost the detection rate of lymph node. In events of lymph node metastasis or no extracapsular invasion, carbon nanoparticles can adequately protect parathyroid functions.

[The role of medical imaging plus carbon nanoparticles to manage the cervical lymph nodes in patients with thyroid carcinoma].

OBJECTIVE: The purpose of this study was to discuss the role of the combination of carbon nanoparticles and medical imaging to manage the cervical lymph nodes in patients with thyroid carcinoma. METHOD: Eighty one patients with thyroid carcinoma that primary treated were divided into two groups: trial group and control group. Carbon nanoparticles were injected into the thyroid gland of trial group patients. Central compartment (level VI) dissection, levels IIl and IV dissection, lateral node (levels II-V) dissection were performed respectively in all the patients on the basis of medical imaging and pathology. Total lymph nodes, metastasis lymph nodes, black stained lymph nodes and black stained metastasis lymph nodes of trial group were counted respectively in different dissection specimens. Total lymph nodes and metastasis lymph nodes of control group were counted respectively in different dissection specimens. Parathyroid glands of thyroid or central compartment dissection specimens were counted in two groups. RESULT: In trial group, rate of staining lymph node was 80.0% in central neck dissection tissue, 54.9% in levels III and IV dissection specimen, 39.1% in lateral node dissection specimen. In central compartment dissection tissue, lymph nodes on average in control group were less than in trial group (3.03 ± 2.07 vs. 4.72 ± 2.97) (P < 0.01). The same was in levels III and lV dissection specimen (5.53 ± 3.78 vs. 10.29 ± 3.36) (P < 0.01). As for lateral node dissection specimen,there was no statistic difference in the two group (13.4 ± 9.67 vs. 14.56 ± 6.28) (P > 0.05). There was no statistic difference between control group and trial group for the metastasis lymph nodes in difference dissection specimens. Parathyroid gland was found in 3 thyroid or central compartment dissection specimens among trial group, which was found in 9 specimens among control group, the difference had statistical significance (P < 0.05). CONCLUSION: During levels III and IV dissection in cN0 patients or central compartment dissection, lymph nodes can be signed well by carbon nanoparticles, which can improve the lymph node detection rate, but can not increase the lymph node detection rate in cN+ patients. Parathyroid gland can be preserved by carbon nanoparticles during the thyroid gland resection and central neck dissection.

[The combination of ultrasound and CT evaluate lymph node metastasis of thyroid papillary carcinoma in different compartments].

OBJECTIVE: To evaluate the role of the combination of ultrasound and enhanced CT in analyzing lymph node metastasis in thyroid papillary carcinoma (PTC) patients by compartment. METHOD: Clinical data of 115 cases (141 sides) with PTC were collected. All had undergone ultrasound in neck and enhanced CT both in neck and in mediastinum before surgery. They were divided into ultrasound group. CT group, and the combination of ultrasound and enhanced CT group to evaluate lymph node metastasis. RESULT: For the central compartment, the accuracy of ultrasound was 61.0%. CT was 48.9%, and the combination of ultrasound and CT was 62.4%. For the lateral compartment, ultrasound was 87.9%, CT was 78.7%, the combination of ultrasound and CT was 85.8%. Ultrasound had higher accuracy than CT in the central (P < 0.05) and lateral (P < 0.05) compartment. The combination of ultrasound and CT had higher accuracy than CT in the central compartment (P < 0.05), but there was no significant difference in the lateral compartment (P > 0.05). There was no significant difference in accuracy between ultrasound and the combination of ultrasound and CT neither in central (P > 0.05) nor in lateral (P > 0.05) compartment. Six cases of lymph node metastasis in mediastinum and 1 case in parapharyngeal space detected by CT were pathologically proven. CT found that five patients with pulmonary metastasis. CONCLUSION: The combination of ultrasound and CT or single ultrasound has higher accuracy in preoperative evaluation than single CT for lymph node metastasis in PTC. CT can assess some compartments such as mediastinum which can't be detected by ultrasound, and at the same time to evaluate lung metastasis. To evaluate lymph node metastasis in PTC, the combination of ultrasound and CT is more accurate and considerate than single method.

[Expression and methylation status of IGFBP-rP1 gene in laryngocarcinoma].

OBJECTIVE: To investigate the relationship between the promoter methylation and protein expression of insulin-like growth factor binding protein-related protein 1(IGFBP-rP1) in laryngeal squamous cell carcinoma (LSCC). METHOD: Methylation specific PCR (MSP) approach and immunohistochemistry methods were used to examine the methylation status and protein expression of IGFBP-rP1 in 45 samples of laryngeal carcinoma and 18 samples of tissues beside cancer. RESULT: For the promoter site, methylation frequency of IGFBP-rP1 in tumor specimens (33.3%, 15/45) was significantly higher than that in corresponding normal tissues (5.6%, 1/18) (P < 0.05). The protein expression of IGFBP-rP1 in tumor tissues was significantly higher than that in corresponding normal tissues (P < 0.05) and was inversely correlated with its methylation status of promoter. CONCLUSION: Epigenetic silencing of IGFBP-rP1 gene expression by promoter hypermethylation may play an important role in LSCC.