COA6 promotes the oncogenesis and progression of breast cancer by oxidative phosphorylation pathway.

Mitochondrial oxidative phosphorylation (OXPHOS) has long been considered the primary energy source in breast cancer cells. Cytochrome c oxidase assembly factor 6 (COA6), which functions as a metal chaperone to transport copper to complex Ⅳ during the OXPHOS process, plays a crucial role in the carcinogenesis of lung adenocarcinoma. Nevertheless, its specific function in breast cancer is undefined. The present investigation aimed to clarify COA6's expression profile and regulatory functions in breast cancer, as well as to unveil its underlying mechanisms. Initially, our findings revealed a significant upregulation of COA6 in breast cancer, as evidenced by an analysis of the TCGA database and tissue microarrays. This upregulation correlated with tumor size and histological grade. Additionally, survival analysis revealed that elevated COA6 amounts were correlated with decreased overall survival (OS) in breast cancer. To delve deeper into the functions of COA6, both COA6-overexpressing and COA6-knockdown breast cancer cell models were established. These experiments demonstrated COA6 is pivotal in regulating cell proliferation, apoptosis, migration, and invasion, thereby promoting cancer progression in vitro. Notably, functional enrichment analysis indicated COA6 might be involved in breast cancer progression by modulating oxidative phosphorylation (OXPHOS). Collectively, this study reveals an overt tumorigenic role for COA6 in breast cancer and sheds light on its potential mechanisms, offering valuable therapeutic targets for breast cancer therapy.

Increased expression of REG3A promotes tumorigenic behavior in triple negative breast cancer cells.

BACKGROUND: Identifying new targets in triple negative breast cancer (TNBC) remains critical. REG3A (regenerating islet-derived protein 3 A), a calcium-dependent lectin protein, was thoroughly investigated for its expression and functions in breast cancer. METHODS: Bioinformatics and local tissue analyses were employed to identify REG3A expression in breast cancer. Genetic techniques were employed to modify REG3A expression, and the resulting effects on the behaviors of breast cancer cells were examined. Subcutaneous xenograft models were established to investigate the involvement of REG3A in the in vivo growth of breast cancer cells. RESULTS: Analysis of the TCGA database uncovered increased REG3A levels in human breast cancer tissues. Additionally, REG3A mRNA and protein levels were elevated in TNBC tissues of locally treated patients, contrasting with low expression in adjacent normal tissues. In primary human TNBC cells REG3A shRNA notably hindered cell proliferation, migration, and invasion while triggering caspase-mediated apoptosis. Similarly, employing CRISPR-sgRNA for REG3A knockout showed significant anti-TNBC cell activity. Conversely, REG3A overexpression bolstered cell proliferation and migration. REG3A proved crucial for activating the Akt-mTOR cascade, as evidenced by decreased Akt-S6K1 phosphorylation upon REG3A silencing or knockout, which was reversed by REG3A overexpression. A constitutively active mutant S473D Akt1 (caAkt1) restored Akt-mTOR activation and counteracted the proliferation inhibition and apoptosis induced by REG3A knockdown in breast cancer cells. Crucially, REG3A played a key role in maintaining mTOR complex integrity. Bioinformatics identified zinc finger protein 680 (ZNF680) as a potential REG3A transcription factor. Knocking down or knocking out ZNF680 reduced REG3A expression, while its overexpression increased it in primary breast cancer cells. Additionally, enhanced binding between ZNF680 protein and the REG3A promoter was observed in breast cancer tissues and cells. In vivo, REG3A shRNA significantly inhibited primary TNBC cell xenograft growth. In REG3A-silenced xenograft tissues, reduced REG3A levels, Akt-mTOR inhibition, and activated apoptosis were evident. CONCLUSION: ZNF680-caused REG3A overexpression drives tumorigenesis in breast cancer possibly by stimulating Akt-mTOR activation, emerging as a promising and innovative cancer target.

[Comparison of results of prenatal diagnosis by different techniques for fetuses with increased nuchal translucency].

OBJECTIVE: To assess the value of chromosomal microarray analysis (CMA) and trio-whole exome sequencing (trio-WES) for fetuses with increased nuchal translucency (NT) thickness. METHODS: Sixty two pregnant women who had visited Urumqi Maternal and Child Care Health Hospital between June 2018 and June 2020 for NT ≥ 3.0 mm at 11 ~ 13+6 gestational weeks were selected as study subjects. Relevant clinical data were collected. The patients were divided into 3.0 ~ ＜3.5 mm (n = 33) and ≥3.5 mm groups (n = 29). Chromosome karyotyping analysis and chromosomal microarray analysis were carried out. And trio-WES analysis was performed on 15 samples with NT thickening but negative CMA results. The distribution and incidence of chromosomal abnormalities in the two groups were compared by using chi-square test. RESULTS: The median age of the pregnant women was 29 years old (22 ~ 41 years old), the median thickness of NT was 3.4 mm (3.0 ~ 9.1 mm), and the median gestational age at the detection was 13+4 weeks (11+5 ~ 13+6 weeks). Chromosome karyotyping analysis has detected 12 cases of aneuploidies and 1 case of derivative chromosome. The detection rate was 20.97% (13/62). CMA has detected 12 cases of aneuploidies, 1 case of pathogenic CNV and 5 cases of variant of uncertain significance (VUS), with a detection rate of 29.03% (18/62). The aneuploidy rate for the NT ≥ 3.5 mm group was higher than that for the 3.0 ≤ NT < 3.5 mm group [3.03% (1/33) vs. 41.38% (12/29), χ² = 13.698, P < 0.001]. There was no statistically significant difference between the two groups in the detection rate of fetal pathogenic CNV and VUS (χ² = 0.028, P > 0.05). Trio-WES analysis of 15 samples with negative CMA result and no structural abnormality has identified 6 heterozygous variants, including SOS1: c.3542C>T (p.A1181V) and c.3817C>G (p.L1273V), COL2A1: c.436C>T (p.P146S) and c.3700G>A (p.D1234N), LZTR1: c.1496T>C (p.V499A), and BRAF: c.64G>A (p.D22N), respectively. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), all of the variants were rated as VUS. CONCLUSION: NT thickening can indicate chromosome abnormality, and CMA and trio-WES may be used for the prenatal diagnosis.

UGT1A1 variants in Chinese Uighur and Han newborns and its correlation with neonatal hyperbilirubinemia.

To explore the correlation between UGT1A1 variant and neonatal hyperbilirubinemia in Chinese Uighur and Han populations. We conducted this study in Urumqi, China. Umbilical cord blood specimens and clinical information of term infants born in the studied center were collected. Variation status of UGT1A1 was determined by direct sequencing or capillary electrophoresis analysis. 102 Uighur and 99 Han normal term neonates, together with 19 hospitalized term newborns (10 Uighur and 9 Han) due to significant hyperbilirubinemia were enrolled into the final analysis. The incidence of neonates with high-risk transcutaneous bilirubin level (TCB) were much higher in Han newborns than in Uighur newborns(P = 0.01). Also, there was statistically significant difference in (TA) 7 promoter mutation of UGT1A1 between Han and Uighur group(χ2 = 4.675, P = 0.03). Furthermore, exon mutation (c.211 and /or c.1091) in UGT1A1 gene was significantly associated with increased TCB level (ORadj = 1.41, 95%CI: 0.25-2.51, P = 0.002) and higher risk of hyperbilirubinemia in both Han and Uighur infants after adjusted for covariates (ORadj = 2.21, 95%CI: 1.09-4.49, P = 0.03). In conclusion, UGT1A1 promoter polymorphism seem to be an important genetic modulator of plasma bilirubin level and neonatal hyperbilirubinemia risk within ethnic groups. Genetic assessment of UGT1A1 coding variants may be useful for clinical diagnosis of neonatal jaundice.

Indocyanine green near-infrared imaging-guided lymph node dissection during oesophageal cancer surgery: A single-centre experience.

Objective: This study aimed to investigate the feasibility of using indocyanine green (ICG) near-infrared (NIR) imaging during lymphadenectomy for oesophageal cancer. Methods: Eighty-seven patients with primary oesophageal cancer were enrolled in this study. All the enrolled patients received an endoscopic injection of ICG between 40 min and 23 h before surgery. Nodal dissection during surgery was performed under fluorescence imaging visualisation, with the NIR signal shown in purple. ICG+ or ICG- nodes were recorded station by station and were microscopically evaluated. Results: Endoscopic peritumoral ICG injection was successfully performed in all patients. Major post-surgery complications included wound infection, pleural effusion, dysphonia, pneumonia and anastomotic fistula. No patients experienced ICG-related adverse events. A total of 2,584 lymph nodes were removed, and the mean number of lymph nodes for each patient was 29.70 ± 9.24. Most of the removed nodes (97.83%) were ICG+, and 3.32% of the ICG+ nodes were metastatic. No metastatic nodes were ICG- or belonged to an ICG- lymph node station. The time from ICG injection to surgery did not affect the number of harvested lymph nodes. Conclusions: The use of ICG-NIR imaging during oesophageal cancer surgery can enhance the visualisation of lymph nodes during surgery. It is a feasible, safe and helpful technique for lymphadenectomy.

Exosomal lncRNA DOCK9-AS2 derived from cancer stem cell-like cells activated Wnt/ß-catenin pathway to aggravate stemness, proliferation, migration, and invasion in papillary thyroid carcinoma.

Exosomal long non-coding RNAs (lncRNAs) are crucial factors that mediate the extracellular communication in tumor microenvironment. DOCK9 antisense RNA2 (DOCK9-AS2) is an exosomal lncRNA which has not been investigated in papillary thyroid carcinoma (PTC). Based on the result of differentially expressed lncRNAs in PTC via bioinformatics databases, we discovered that DOCK9-AS2 was upregulated in PTC, and presented elevation in plasma exosomes of PTC patients. Functionally, DOCK9-AS2 knockdown reduced proliferation, migration, invasion, epithelial-to-mesenchymal (EMT) and stemness in PTC cells. PTC-CSCs transmitted exosomal DOCK9-AS2 to improve stemness of PTC cells. Mechanistically, DOCK9-AS2 interacted with SP1 to induce catenin beta 1 (CTNNB1) transcription and sponged microRNA-1972 (miR-1972) to upregulate CTNNB1, thereby activating Wnt/ß-catenin pathway in PTC cells. In conclusion, PTC-CSCs-derived exosomal lncRNA DOCK9-AS2 activated Wnt/ß-catenin pathway to aggravate PTC progression, indicating that DOCK9-AS2 was a potential target for therapies in PTC.

Elevated O-GlcNAcylation Promotes Malignant Phenotypes of Hypopharyngeal Squamous Cell Carcinoma by Stabilizing Nrf2 through Regulation of the PI3K/Akt Pathway.

BACKGROUND AND PURPOSE: O-GlcNAcylation is a significant protein posttranslational modification with O-linked ß-N-acetylglucosamine (GlcNAc) for intracellular signaling. Elevated O-GlcNAcylation contributes to cell proliferation, cell migration, cell apoptosis and signal transduction in various cancers. However, the expression level and functional role of O-GlcNAcylation in Hypopharyngeal Squamous Cell Carcinoma (HSCC) is not clearly elucidated. Nuclear factor erythroid-2-related factor 2 (Nrf2) is a master transcriptional factor that has been found to be aberrantly activated in HSCC. Here, we provide a molecular rationale between O-GlcNAcylation and Nrf2 in HSCC patients. METHODS: The protein levels of O-GlcNAcylation and Nrf2 in HSCC tissues were detected by immunohistochemistry technique and western blot analysis. Then, O-GlcNAcylation knockdown HSCC cells were applied in this study. Cell proliferation was detected by CCK8, colony-forming analysis, and cell cycle assays. Cell migration and invasion ability was evaluated by transwell assays. Cell apoptosis was measured by TUNEL analysis. RESULTS: O-GlcNAcylation was obviously up-regulated in HSCC tissues, which correlated with tumor size and lymph node metastasis. In addition, the protein level of Nrf2 was found to positively correlate with the expression of O-GlcNAcylation both in vivo and in vitro. Knockdown of O-GlcNAcylation significantly inhibited HSCC cell growth, suppressed cell migration, and promoted cell apoptosis, whereas overexpression of Nrf2 reversed these phenotypes. Mechanismly, the upregulation of O-GlcNAcylation promoted the phosphorylation of Akt, leading to the stabilization of Nrf2; this could be attenuated by inhibition of the PI3K/Akt signaling pathway. CONCLUSION: Here, we provide a molecular association between O-GlcNAcylation and Nrf2 in HSCC patients, thus providing valuable therapeutic targets for the disease.

Transcriptome analysis of tongue cancer based on high­throughput sequencing.

Tongue cancer is one of the most common types of cancer, but its molecular etiology and pathogenesis remain unclear. The aim of the present study was to elucidate the pathogenesis of tongue cancer and investigate novel potential diagnostic and therapeutic targets. Four matched pairs of tongue cancer and paracancerous tissues were collected for RNA sequencing (RNA­Seq), and the differentially expressed genes were analyzed. The RNA­Seq data of tongue cancer tissues were further analyzed using bioinformatics and reverse transcription­quantitative PCR analysis. The sequenced reads were quantified and qualified in accordance with the analysis demands. The transcriptomes of the tongue cancer tissues and paired paracancerous tissues were analyzed, and 1,700 upregulated and 2,249 downregulated genes were identified. Gene Ontology analysis uncovered a significant enrichment in the terms associated with extracellular matrix (ECM) organization, cell adhesion and collagen catabolic processes. Kyoto Encyclopedia of Genes and Genomes analysis demonstrated that these differentially expressed genes were mainly enriched in the focal adhesion pathway, ECM­receptor interaction pathway, phosphoinositide 3­kinase (PI3K)­Akt pathway, and cell adhesion molecules. Comprehensive analyses of the gene tree and pathway network revealed that the majority of cell cycle genes were upregulated, while the majority of the genes associated with intracellular response, cell adhesion and cell differentiation were downregulated. The ECM­receptor interaction, focal adhesion kinase (FAK) and PI3K­Akt pathways were closely associated with one another and held key positions in differential signaling pathways. The ECM­receptor, FAK and PI3K­Akt signaling pathways were found to synergistically promote tongue cancer occurrence and progression, and may serve as potential diagnostic and therapeutic targets for this type of cancer.

[Application for prenatal diagnosis using both chromosomal karyotype analysis and BACs-on-Beads assay].

OBJECTIVE: To assess the application value in prenatal diagnosis using karyotype analysis combined with BACs-on-Beads (BoBs) assay. METHODS: Nine hundred sixty five pregnant women were subjected to amniocentesis, chromosomal karyotype analysis and detection of BoBs were employed simultaneously for abnormal number of chromosomes and 9 chromosome microdeletion syndrome in prenatal diagnosis. RESULTS: Fifty cases common chromosome aneupoidies were successfully detected by both karyotype analysis and BoBs which included 31 cases of trisomy 21,10 cases of trisomy 18 and 9 cases with sex chromosome abnormality. BoBs in addition detected 1 case of DiGeorge-1 microdeletion syndrome and 1 case of 7q11.23 microduplication syndrome. All 9 fetuses with chromosome abnormalities detected by karyotyping were missed by BoBs, including 2 cases of marker chromosomes,4 cases of chromosomal translocation,1 case of chromosomal inversion, 2 cases of Sex chromosome mosaicism; 2 cases of fetal inherited from the parents,7 cases for novel mutations. CONCLUSION: Karyotype analysis combined with BoBs dedtection is a rapid, effective and highly accurate prenatal diagnosis model that may should be widely used in clinical diagnosis.

Down-regulation of long non-coding RNA AFAP1-AS1 inhibits tumor growth, promotes apoptosis and decreases metastasis in thyroid cancer.

Long non-coding RNAs (lncRNAs), a new type of transcripts, play important roles in various cellular biological processes, involving tumorigenesis. Previous studies showed that lncRNA AFAP1-AS1 was aberrantly expressed in numerous cancers. Nevertheless, we know quite a little about the expression pattern and biological function of AFAP1-AS1 in thyroid cancer. In this study, we adopted the quantitative real-time PCR (qRT-PCR) to detect the expression of AFAP1-AS1 in thyroid cancer tissues. We discovered that expression of AFAP1-AS1 was increased in thyroid cancer tissues. MTT assays elucidated that down-regulation of AFAP1-AS1 could suppress growth of thyroid cancer cells. And the results of flow cytometry analysis indicated knockdown of AFAP1-AS1 induced apoptosis in thyroid cancer. Transwell assay was applied to show decreased cell migration in thyroid cancer as a result of down-regulation of AFAP1-AS1. Hence, our study provided evidence for our hypothesis that AFAP1-AS1 could be a therapeutic target for thyroid cancer.

[Prenatal diagnosis of a rare case of 7q11.23 duplication syndrome].

OBJECTIVE: To explore the application of combined techniques for the prenatal diagnosis of a case with 7q11.23 duplication. METHODS: Amniocentesis was performed in the second trimester for a mother with a high risk suggested by serological prenatal screening. G-banded chromosomal analysis was performed on cultured amniocytes and peripheral blood samples from both parents. DNA from amniotic fluid sample was isolated for a BACs-on-Beads (BoBs) assay. To define the range of duplication, copy number variation was determined with single nucleotide polymorphism array (SNP array, Affymetrix CytoScan 750K) and fluorescence in situ hybridization (FISH) analysis. RESULTS: Chromosomal analysis suggested that the fetus and both parents all had a normal karyotype, while a duplication of 7q11.23 was detected by the BoBs assay. SNP array revealed a 1.5 Mb duplication in chromosome 7q11.23, which was confirmed by FISH. CONCLUSION: Combined prenatal BoBs, SNP array and FISH has enabled effective diagnose of a case with 7q11.23 syndrome.

[The significance of lymph node dissection in the VI area of cN0 thyroid papillary carcinoma].

OBJECTIVE: The significance of lymph node dissection in the VI area of cN0 thyroid papillary carcinoma. METHOD: Collect 150 cases of patients diagnosed with cNO thyroid papillary carcinoma and they were performed thyroid gland lobe and isthmic portion excision including lateral VI area lymph node cleaning. The specimens were pathologic examined to determinate the size, the position, invasion of thyroid papillary carcinoma,the number and metastasis of lymph node, etc. RESULT: In the 150 patients performed the lymph node VI area groups cleaning, 93 cases had VI area of lymph node metastases, so the transfer rate was 62.0%. In the VI area, metastasis rate of tracheal side lymph nodes was 62.0% (93/150), lymph node before throat group was 4.67% (7/150), lymph node before trachea group was 3.33% (5/150), lymph nodes near the trachea laryngeal recurrent nerve ventral group was 52.0% (78/150), and next to the trachea laryngeal recurrent nerve dorsal lymph node group was 21.33% (32/ 150). CONCLUSION: In CN0 thyroid papillary carcinoma, VI zone of lymph node metastasis rate is high, and region VI lymph node metastasis rate from high to low in order for: paratracheal lymph node, prelaryngeal lymph node, pretracheal lymph node. The metastasis rate of paratracheal throat back nerve ventral lymph node was the highest in central lymph node.

[Relationship between genetic polymorphisms of DAT1 VNTR and cancer with abnormal Hilit].

OBJECTIVE: This study was to investigate the relationship between the 40 bp VNTR polymorphism of dopamine transporter gene (DAT1) and cancer with Uighur medicine abnormal Hilit on Chinese Uighur population of Xinjiang province. METHODS: Divided the cancer patients into four body fluids according to Uighur medicine theory, the polymerase chain reaction and VNTR polymorphism technique was employed to detect genotype and allele frequencies of a 40 bp VNTR polymorphism situated in 3' untranslated region of the DAT1 gene in 47 cancer patients with abnormal black Savda, 26 cancer patients with other abnormal Hilit and 57 normal control subjects in Uighur population of Xinjiang Province. RESULTS: (1) In our sample,the repeat numbers of 40 bp were 6 to 11 (PCR product length of 320 bp to 520 bp) and 10-repeats allele (480 bp) detected was the dominant allele of DAT1 gene polymorphisms with the allele frequency 90.4%. Six kinds of genotype were detected in this study and the genotype 480 bp/480 bp was the most common genotype with genotype frequency 80.7%. (2) The susceptibility to cancer patients with abnormal black Savda among the subjects with 10 repeat (R) allele and 10/10 genotypes and the subjects with non-10 repeat (R) allele and non-10/10 genotypes were similar (P = 0.158, OR = 1.994, 95% CI = 0.754-5.275; P = 0.138, OR = 2.143, 95% CI = 0.772-5.947). No significant differences for the genotype frequency or the allele frequency of the 40 bp VNTR polymorphism of DAT1 were revealed between cancer patients with abnormal black Savda and cancer patients with other abnormal Hilit (P = 0.729, P = 0.782). CONCLUSION: The 40 bp VNTR polymorphism of DAT1 may not be correlated to the susceptibility to cancer with Uighur medicine abnormal Hilit.

[Association analysis between polymorphism of dopamine transporter variable number tandem repeat and breast cancer with abnormal Hilit].

OBJECTIVE: To explore the association between polymorphism of dopamine 1 transporter variable number tandem repeat (DAT1 VNTR) and breast cancer with abnormal Hilit in Chinese Han population from Xinjiang. METHODS: The breast caner patients were divided into four body fluids according to Uighur medicine theory. And polymerase chain reaction and VNTR polymorphism technique were employed to detect genotypic and allelic frequencies of a 40 bp VNTR polymorphism situated in 3' untranslated region of DAT1 gene in 144 breast cancer patients with abnormal Hilit and 104 normal control subjects in Han population of Xinjiang province. RESULTS: In our sample, the repeat numbers of 40 bp were 7 and 9 - 11 (PCR product length of 360 bp and 440 bp to 520 bp) and 10-repeat allele (480 bp) detected was the dominant allele of DAT1 gene polymorphisms with an allelic frequency of 90.9%; Seven kinds of genotype were detected and genotype 10/10 was the most common genotype with a genotypic frequency of 83.1%. The frequency of 10-repeats allele and 10/10 genotype was significantly higher in breast caner patients with abnormal balgham Hilit than in controls (OR = 0.127, 95%CI 0.016 - 0.988, P = 0.026; OR = 0.134, 95%CI 0.018 - 1.016, P = 0.020) and breast caner patients with abnormal Sapra Hilit (OR = 0.132, 95%CI 0.016 - 1.075, P = 0.049; OR = 0.132, 95%CI 0.017 - 1.042, P = 0.033). CONCLUSION: The 10-repeats allele and 10/10 genotype of 40 bp VNTR polymorphism of DAT1 may increase the risk of breast cancer with Uighur medicine abnormal Hilit in Chinese Han population from Xinjiang province and it is not correlated with the susceptibility to breast cancer with Uighur medicine abnormal Sapra Hilit.

[Study of serotonin transporter gene polymorphisms in disease of patients with abnormal black Savda].

OBJECTIVE: To explore the association between patients with abnormal black Savda according to Uighur medicine theory and the serotonin transporter (5-HTT) gene polymorphisms in the promoter (5-HTTLPR) and the variable number tandem repeats (VNTRs) in intron 2 (5-HTTVNTR). METHODS: Divided the patients into four body fluids according to Uighur medicine theory, PCR was used to detect genotype and allele frequencies of 5-HTILPR and 5-HTTVNTR polymorphisms in 66 patients with abnormal black Savda (including 35 cases of hypertension, 19 cases of diabetes, 12 cases of cancer diagnosis) and 45 control subjects. RESULTS: No significant differences for the genotype frequency or the allele frequency of 5-HTTLPR and 5-HTTVNTR were revealed between patients with abnormal black Savda and control subjects. More control subjects tended to be of L/L genotype (15.56%) than hypertension with abnormal black Savda (5.72%). On the contrary, more cancer diagnosis with abnormal black Savda tended to be of L/L genotype (25.00%) and L (54.17%) allele than control subjects L/L genotype (15.56%) and L allele (37.78%). On the other hand, fewer control subjects tended to be of 10/10 genotype (4.45%) than diabetes with abnormal black Savda (10.53%) and cancer diagnosis with abnormal black Savda (8.34%). CONCLUSION: 5-HTTLPR L/L genotype may be a protective factor for hypertension with abnormal black Savda, 5-HTTLPR L/L genotype and L allele may be risk factors for cancer diagnosis with abnormal black Savda, 5-HTTVNTR 10/10 genotype may be risk factors for cancer diagnosis with abnormal black Savda and diabetes with abnormal black Savda.