OBJECTIVES: To explore the molecular mechanism by which lncRNA SNHG15 regulates proliferation, invasion and migration of lung adenocarcinoma cells. METHODS: The lncRNA microarray chip dataset GSE196584 and LncBase were used to predict the lncRNAs that interact with miR-30b-3p, and their association with patient prognosis were investigated using online databases, after which lncRNA nucleolar RNA host gene 15 (SNHG15) was selected for further analysis. The subcellular localization of lncRNA SNHG15 and its expression levels in normal human lung epithelial cells and lung adenocarcinoma cell lines were detected using fluorescence in situ hybridization and qRT-PCR. In cultured A549 cells, the changes in cell proliferation, migration, and invasion following transfection with a SNHG15 knockdown plasmid (sh-SNHG15), a miR-30b-3p inhibitor, or their co-transfection were assessed with EdU, wound healing, and Transwell assays. Bioinformatics analyses were used to predict the regulatory relationship between lncRNA SNHG15 and COX6B1, and the results were verified using Western blotting and rescue experiments in A549 cells transfected with sh-SNHG15, a COX6B1-overexpressing plasmid, or both. RESULTS: LncRNA SNHG15 was shown to target miR-30b-3p, and the former was highly expressed in lung adenocarcinoma, and associated with a poor patient prognosis. LncRNA SNHG15 was localized in the cytoplasm and expressed at higher levels in A549 and NCI-H1299 cells than in BEAS-2B cells. In A549 cells, lncRNA SNHG15 knockdown significantly inhibited cell migration, invasion and proliferation, and these changes were reversed by miR-30b-3p inhibitor. A regulatory relationship was found between lncRNA SNHG15 and COX6B1, and their expression levels were positively correlated (r=0.128, P=0.003). MiR-30b-3p knockdown obviously decreased COX6B1 expression in A549 cells, and COX6B1 overexpression rescued the cells from the inhibitory effects of lncRNA-SNHG15 knockdown. CONCLUSIONS LncRNA SNHG15 may compete with COX6B1 to bind miR-30b-3p through a ceRNA mechanism to affect proliferation, migration, and invasion of lung adenocarcinoma cells.
Humans ; MicroRNAs/metabolism* ; RNA, Long Noncoding/genetics* ; Cell Proliferation ; Cell Movement ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung ; Neoplasm Invasiveness ; A549 Cells ; Adenocarcinoma/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Line, Tumor

Objective:To study the effects of Linc00426 on the migration,invasion and proliferation of oral squamous carcinoma cells(OSCC).Methods:TCGA database was used to query the differential expression of Linc00426.RT-qPCR was used to detect the expression level of Linc00426 in normal human oral keratinocytes(NHOK)cells and various OSCC cells lines,the knockout effi-ciency of Linc00426 at 3 sites and the expression level of miR-455-5p.Transwell test and EdU test were used to detect the changes of the migration,invasion and proliferation of CAL27 cells.Bioinformatics databases was used to predict subcellular localization and downstream miRNAs and target proteins of Linc00426.Western blot assay was used to detect the changes of USP3 expression levels in CAL27 cells.Results:The expression level of Linc00426 was increased in OSCC tissue(P＜0.05),and its expression in CAL27,HN4 and HN30 cells was higher than that in NHOK cells(P＜0.05).Knocking out Linc00426 inhibited the migration,invasion,and proliferation ability of CAL27 cells(P＜0.05).The bioinformatics database indicates that the subcellular localization of Linc00426 is in the cytoplasm;Linc00426 is negatively correlated with miR-455-5p,and miR-455-5p is negatively correlated with USP3,in which there are targeted binding sites.Knocking out miR-455-5p promoted the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knocking out USP3 inhibited the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knock out Linc00426 reduced the expression level of USP3,while knocking out miR-455-5p increased the expression level of USP3(P＜0.05).Conclusion:Linc00426 plays a role as an oncogene in OSCC cells and could inhibit the migration,invasion and proliferation of the cells by targeting USP3 to regulate miR-455-5p.

Objective To investigate the expression levels and clinical significance of serum long chain non coding RNA cancer susceptibility candidate 11(lncRNA CASC11)and Testis-associated highly-conserved oncogenic long non-coding RNA(lncRNA THOR)in esophageal cancer patients.Methods Esophageal cancer patients diagnosed in our hospital from January 2017 to January 2021 were selected as the observation group(n=129),and healthy individuals who underwent physical examination in our hospital(n=129)were as the control group.Serum lncRNA CASC11 and lncRNA THOR expression levels were detected by RT-qPCR;the 3-year survival rate was analyzed by Kaplan-Meier method;and the diagnostic value was analyzed by ROC curve.Results The expression levels of serum lncRNA CASC11(1.48±0.35 vs 1.01±0.20)and lncRNA THOR(1.54±0.28 vs 1.11±0.15)in the observation group were obviously higher than those in the control group(P＜0.05).The proportions of high expression of serum lncRNA CASC11 and lncRNA THOR in patients with infiltration depth of T3～T4,tumor diameter＞4 cm,TNM staging of Ⅲ-Ⅳ,and lymph node metastasis were obviously higher than those in patients with infiltration depth T1～T2,tumor diameter ≤ 4 cm,TNM staging Ⅰ-Ⅱ,and no lymph node metastasis(P＜0.05).After 3-year of follow-up,52 out of 129 esophageal cancer patients died and 77 survived.The 3-year survival rate of patients with high expression of lncRNA CASC11(32.20％vs 82.86％)and lncRNA THOR(36.07％vs 80.88％)was obviously lower than that of patients with low expression of lncRNA CASC11 and lncRNA THOR(P＜0.05).The area under the curve(AUC)of serum lncRNA CASC11 and lncRNA THOR levels for diagnosing esophageal cancer was 0.888 and 0.914,respectively,with cutoff value of 1.29 and 1.32.The AUC of the combined diagnosis of the two was 0.946,and the combined diagnosis of the two was superior to their individual diagnosis(Zcombination-lncRNA CASC11=2.410,Zcombination-lncRNA THOR=2.167,P=0.010,0.040).Conclusion The serum levels of lncRNA CASC11 and lncRNA THOR in patients with esophageal cancer are upregulated,they are obviously associated with the 3-year survival rate of patients,and the combination of the two has higher efficacy in the diagnosis of esophageal cancer.

Objective To investigate the expression levels and clinical significance of serum long chain non coding RNA cancer susceptibility candidate 11(lncRNA CASC11)and Testis-associated highly-conserved oncogenic long non-coding RNA(lncRNA THOR)in esophageal cancer patients.Methods Esophageal cancer patients diagnosed in our hospital from January 2017 to January 2021 were selected as the observation group(n=129),and healthy individuals who underwent physical examination in our hospital(n=129)were as the control group.Serum lncRNA CASC11 and lncRNA THOR expression levels were detected by RT-qPCR;the 3-year survival rate was analyzed by Kaplan-Meier method;and the diagnostic value was analyzed by ROC curve.Results The expression levels of serum lncRNA CASC11(1.48±0.35 vs 1.01±0.20)and lncRNA THOR(1.54±0.28 vs 1.11±0.15)in the observation group were obviously higher than those in the control group(P＜0.05).The proportions of high expression of serum lncRNA CASC11 and lncRNA THOR in patients with infiltration depth of T3～T4,tumor diameter＞4 cm,TNM staging of Ⅲ-Ⅳ,and lymph node metastasis were obviously higher than those in patients with infiltration depth T1～T2,tumor diameter ≤ 4 cm,TNM staging Ⅰ-Ⅱ,and no lymph node metastasis(P＜0.05).After 3-year of follow-up,52 out of 129 esophageal cancer patients died and 77 survived.The 3-year survival rate of patients with high expression of lncRNA CASC11(32.20％vs 82.86％)and lncRNA THOR(36.07％vs 80.88％)was obviously lower than that of patients with low expression of lncRNA CASC11 and lncRNA THOR(P＜0.05).The area under the curve(AUC)of serum lncRNA CASC11 and lncRNA THOR levels for diagnosing esophageal cancer was 0.888 and 0.914,respectively,with cutoff value of 1.29 and 1.32.The AUC of the combined diagnosis of the two was 0.946,and the combined diagnosis of the two was superior to their individual diagnosis(Zcombination-lncRNA CASC11=2.410,Zcombination-lncRNA THOR=2.167,P=0.010,0.040).Conclusion The serum levels of lncRNA CASC11 and lncRNA THOR in patients with esophageal cancer are upregulated,they are obviously associated with the 3-year survival rate of patients,and the combination of the two has higher efficacy in the diagnosis of esophageal cancer.

Objective:To investigate the clinicopathological characteristics of ossifying fibromyxoid tumor (OFMT) with rare fusion subtypes.Methods:Three cases of OFMT with rare fusion subtypes, diagnosed and consulted in the Zhejiang Hospital, Zhejiang Provincial People′s Hospital, Hangzhou, China and Ningbo Clinical Pathology Diagnosis Center, Ningbo, China from January 2016 to December 2024 were collected. Immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and targeted RNA sequencing were performed to analyze the immunohistochemical and molecular genetic characteristics of these OFMT. Literature review was also conducted.Results:All three patients were male, with ages of 50, 74, and 58 years, respectively. The tumors were located in the left foot, left thigh, and left lumbar region, respectively, and all presented as slowly growing, painless masses in the skin or subcutaneous tissue. Grossly, the tumors measured 3.5 cm, 6.3 cm, and 5.0 cm in maximum diameter, respectively, with a grayish-white to grayish-yellow, solid, lobulated cut surface. One case exhibited a noticeable myxoid texture. Microscopically, one tumor was located in the superficial dermis, while the other two were in the subcutaneous tissue. The tumors were well-demarcated and showed a lobulated or multinodular growth pattern. None of the cases had a complete surrounding bony shell (only one case had very focal ossification). The tumor cells were monomorphic, short spindle-shaped, oval to epithelioid, and arranged in solid sheets, trabeculae, and small nests within a variably fibromyxoid stroma. Case 1 exhibited abundant pseudorosette-like structures formed by short spindle cells surrounding acellular fibrous stroma. Case 2 showed focal transition of epithelioid tumor cells into fasciculately arranged spindle cells, with extensive stromal hyalinization. Case 3 had a predominantly myxoid stroma with a rich network of thin-walled blood vessels. The tumor cells exhibited mild nuclear atypia with 1-3 mitotic figures per 50 high-power fields. All three cases showed diffuse and strong expression of CD10. Two of the three cases showed nuclear expression of TFE3, while one case showed diffuse and strong expression of desmin and S-100. Targeted RNA sequencing revealed PHF1 (ex12)::TFE3 (ex7) fusion in two cases and MEAF6 (ex5)::PHF1 (5′UTR) fusion in one case, which were further confirmed by FISH study. All three patients underwent tumor resection. Two showed no recurrence during follow-up periods of 98 months and 15 months, respectively, while one experienced local recurrence at 12 months postoperatively.Conclusions:OFMT with rare fusion subtypes often exhibits atypical histological and immunophenotypic features, and lacks a characteristic bony shell. Incorporating TFE3 into the diagnostic IHC panel greatly aids in screening for the cases with rare PHF1::TFE3 fusions. Familiarity with the histological and immunophenotypic characteristics, and differential diagnostic points of these rare OFMT subtypes, is essential for judicious use of molecular genetic tools in achieving a definitive diagnosis.

Objective:To study the effects of Linc00426 on the migration,invasion and proliferation of oral squamous carcinoma cells(OSCC).Methods:TCGA database was used to query the differential expression of Linc00426.RT-qPCR was used to detect the expression level of Linc00426 in normal human oral keratinocytes(NHOK)cells and various OSCC cells lines,the knockout effi-ciency of Linc00426 at 3 sites and the expression level of miR-455-5p.Transwell test and EdU test were used to detect the changes of the migration,invasion and proliferation of CAL27 cells.Bioinformatics databases was used to predict subcellular localization and downstream miRNAs and target proteins of Linc00426.Western blot assay was used to detect the changes of USP3 expression levels in CAL27 cells.Results:The expression level of Linc00426 was increased in OSCC tissue(P＜0.05),and its expression in CAL27,HN4 and HN30 cells was higher than that in NHOK cells(P＜0.05).Knocking out Linc00426 inhibited the migration,invasion,and proliferation ability of CAL27 cells(P＜0.05).The bioinformatics database indicates that the subcellular localization of Linc00426 is in the cytoplasm;Linc00426 is negatively correlated with miR-455-5p,and miR-455-5p is negatively correlated with USP3,in which there are targeted binding sites.Knocking out miR-455-5p promoted the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knocking out USP3 inhibited the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knock out Linc00426 reduced the expression level of USP3,while knocking out miR-455-5p increased the expression level of USP3(P＜0.05).Conclusion:Linc00426 plays a role as an oncogene in OSCC cells and could inhibit the migration,invasion and proliferation of the cells by targeting USP3 to regulate miR-455-5p.

Objective:To investigate the clinicopathological characteristics of ossifying fibromyxoid tumor (OFMT) with rare fusion subtypes.Methods:Three cases of OFMT with rare fusion subtypes, diagnosed and consulted in the Zhejiang Hospital, Zhejiang Provincial People′s Hospital, Hangzhou, China and Ningbo Clinical Pathology Diagnosis Center, Ningbo, China from January 2016 to December 2024 were collected. Immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and targeted RNA sequencing were performed to analyze the immunohistochemical and molecular genetic characteristics of these OFMT. Literature review was also conducted.Results:All three patients were male, with ages of 50, 74, and 58 years, respectively. The tumors were located in the left foot, left thigh, and left lumbar region, respectively, and all presented as slowly growing, painless masses in the skin or subcutaneous tissue. Grossly, the tumors measured 3.5 cm, 6.3 cm, and 5.0 cm in maximum diameter, respectively, with a grayish-white to grayish-yellow, solid, lobulated cut surface. One case exhibited a noticeable myxoid texture. Microscopically, one tumor was located in the superficial dermis, while the other two were in the subcutaneous tissue. The tumors were well-demarcated and showed a lobulated or multinodular growth pattern. None of the cases had a complete surrounding bony shell (only one case had very focal ossification). The tumor cells were monomorphic, short spindle-shaped, oval to epithelioid, and arranged in solid sheets, trabeculae, and small nests within a variably fibromyxoid stroma. Case 1 exhibited abundant pseudorosette-like structures formed by short spindle cells surrounding acellular fibrous stroma. Case 2 showed focal transition of epithelioid tumor cells into fasciculately arranged spindle cells, with extensive stromal hyalinization. Case 3 had a predominantly myxoid stroma with a rich network of thin-walled blood vessels. The tumor cells exhibited mild nuclear atypia with 1-3 mitotic figures per 50 high-power fields. All three cases showed diffuse and strong expression of CD10. Two of the three cases showed nuclear expression of TFE3, while one case showed diffuse and strong expression of desmin and S-100. Targeted RNA sequencing revealed PHF1 (ex12)::TFE3 (ex7) fusion in two cases and MEAF6 (ex5)::PHF1 (5′UTR) fusion in one case, which were further confirmed by FISH study. All three patients underwent tumor resection. Two showed no recurrence during follow-up periods of 98 months and 15 months, respectively, while one experienced local recurrence at 12 months postoperatively.Conclusions:OFMT with rare fusion subtypes often exhibits atypical histological and immunophenotypic features, and lacks a characteristic bony shell. Incorporating TFE3 into the diagnostic IHC panel greatly aids in screening for the cases with rare PHF1::TFE3 fusions. Familiarity with the histological and immunophenotypic characteristics, and differential diagnostic points of these rare OFMT subtypes, is essential for judicious use of molecular genetic tools in achieving a definitive diagnosis.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most common type of non-small cell lung cancer, and any change of miRNAs expression will affect the degree of target regulation, thus affecting intracellular homeostasis. This study verified that miR-186-5p could inhibit the proliferation, migration and invasion of LUAD cells by regulating PRKAA2. METHODS: Previous investigations found that the expression of miR-186-5p was markedly suppressed in LUAD. Bioinformatics method is used to predict the target protein related to ferroptosis downstream and inquire about its expression level in LUAD and its influence on the survival of patients. Double luciferase verified the binding site of PRKAA2 and miR-186-5p. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression of PRKAA2. The effects of miR-186-5p of LUAD cells as well as the mechanism by which miR-186-5p inhibits Fer-1's sensitivity to ferroptosis were confirmed by EdU, Transwell, and scratch assays. The effect of miR-186-5p on the amount of reactive oxygen species (ROS) in LUAD cells was discovered using ROS experiment. Malondialdehyde (MDA) and glutathione (GSH) experiments were used to detect the effects of miR-186-5p and PRKAA2 on ferroptosis index of LUAD cells. The concentration of lipid ROS (L-ROS) in LUAD cells were measured using the L-ROS tests to determine the effects of miR-186-5p and PRKAA2. RESULTS: The expression of PRKAA2 is up-regulated, and a high level of PRKAA2 expression was associated with a poor prognosis for patients with LUAD. Overexpression of miR-186-5p decreased the gene and protein expression of PRKAA2. By promoting ferroptosis, miR-186-5p overexpression prevented lung cancer cells from proliferating, invading, and migrating. ROS could be produced in higher amounts in LUAD cells due to miR-186-5p. Overexpression of miR-186-5p and knockdown PRKAA2 up-regulated MDA content and reduced GSH content in LUAD cells, respectively. miR-186-5p could increase the content of L-ROS and promote the ferroptosis sensitivity of LUAD cells by targeting PRKAA2. CONCLUSIONS miR-186-5p promotes ferroptosis of LUAD cells through targeted regulation of PRKAA2, thus inhibiting the proliferation, invasion and migration of LUAD.
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Humans ; Lung Neoplasms/genetics* ; Carcinoma, Non-Small-Cell Lung ; Ferroptosis/genetics* ; Reactive Oxygen Species ; Adenocarcinoma of Lung/genetics* ; MicroRNAs/genetics* ; 3,4-Methylenedioxyamphetamine ; Cell Proliferation/genetics* ; Cell Movement/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Line, Tumor ; AMP-Activated Protein Kinases

Objective:To analysis the policy tools and targets of the policies of free training for order-oriented medical students in rural areas of China, for reference for further improving the free medical student training policy.Methods:The research team searched the official websites of the State Council, National Health Commission, Ministry of Education, and other ministries, as well as the Peking University Treasure Database, for national level policy documents related to free training of order-oriented medical students released from June 2010 to May 2023. Policy tool-policy target analysis framework was used to quantitative analysis the policy documents.Results:A total of 16 policy documents were included and 213 policy provisions were extracted. From the perspective of policy tools, the proportion of policy provisions using imperative policy tools was the highest, accounting for 63.38% (135 articles), followed by advisory policy tools(18.78%, 40 articles)and reward based policy tools(13.61%, 29 articles), while functional expansion tools(2.82%, 6 articles) and authoritative restructuring tools(1.41%, 3 articles) accounted for a relatively low proportion. The institutional education stage is the main policy target, with provisions accounting for 76.06% (162 articles), followed by the continuing education stage and the post graduation education stage, accounting for 17.84% (38 articles) and 7.51% (16 articles), respectively. It was uneven distribution of various policy tools and their sub tools within the same policy target.Conclusions:The distribution of policy tools for the free training policy of rural order oriented medical students in China needed to be further balanced. The policy targets were mainly concentrated in the education stage of universities.

Objective:To explore the relationship between executive function and gait in cases of mild amnestic cognitive impairment (aMCI).Methods:Twenty aMCI hospital patients formed an observation group, while 20 healthy counterparts were the control group. Both groups underwent the Tinetti test, followed by the " normal walking" single-task test and the " normal walking + Go/No-go" dual-task test. The pace, step width, stride length, Go/No-go task response time and accuracy rate were recorded.Results:In the single-task test, there was no significant difference in pace or stride width between the two groups, but the average stride length of the observation group (1.11±0.04)cm was significantly shorter than that of the control group. However, in the dual-task test, the average pace time (0.96±0.08)sec and stride length (1.02±0.06)cm of the observation group were significantly smaller than the control group′s averages, while their step width (0.11±0.02)cm was significantly wider. There was no significant difference between the two groups in the response time in a single (Go/No-go) task, but in the dual-task test, the observation group′s average time was significantly longer than the control group′s and the accuracy was significantly poorer. Both the error rate and the non-response rate were significantly higher than among the control group.Conclusions:Mild amnestic cognitive impairment reduces stride length and pace when walking and impairs executive function.