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Objective To investigate the expression of hsa_circ_0018574 in colorectal cancer tissues and human colon cancer HT29 cell line, as well as its effect on the proliferation and apoptosis of colorectal cancer cells. Methods The circPrimer 1.2 software was used to draw the circRNA sequence structure. Meanwhile, the circRNA microarray was used to screen differentially-expressed circRNA in colorectal cancer tissues and adjacent tissues, and RNA was extracted from tissue samples. The expression of hsa_circ_0018574 in human colorectal tumors was detected by RT-qPCR. The si-circ_0018574 was transfected into HT29 cells, and the expression of CDK2, CDK4, CDK6, cyclinD1, and cyclinE cyclins were detected by colony formation assay, flow cytometry, and Western blot, respectively. Results The expression of hsa_circ_0018574 in human colorectal tumor tissues was significantly higher than that in adjacent tissues (P < 0.01). Meanwhile, the si-circ_0018574 in HT29 cells could significantly inhibit the proliferation of cancer cells, reduce clone formation and colony formation ability (P < 0.01), and induce tumor cell apoptosis (P < 0.01). The expressions of CDK2, CDK4, CDK6, cyclinD1 and cyclinE cyclins were decreased. Conclusion The hsa_circ_0018574 is highly expressed in colorectal tumors, and si-circ_0018574 could significantly inhibit the proliferation of HT29 cells, reduce cell division, and induce apoptosis.
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Objective To screen out the biomarkers with diagnostic value in lung cancer by biochip technology. Methods We screened four pairs of differentially-expressed circRNAs in lung cancer by circRNA expression profiling chip, and then verified the screened differential circRNA hsa_circ_0044569 by qRT-PCR, and collected clinical case information of patients at the same time. The independent sample t test and ROC curve were used to analyze the relation between the clinical data of lung cancer patients and the expression of circRNA hsa_circ_0044569 on clinical samples. Results The expression level of hsa_circ_0044569 in lung cancer tissues was higher than that of its paired adjacent tissues. The cutoff value of hsa_circ_0044569 for the diagnosis of lung cancer was 9.62, AUC was 0.758, P < 0.05, sensitivity was 0.712, and specificity was 0.712. The expression of hsa_circ_0044569 was significantly related with the pathological type of lung cancer patients (P < 0.05). Conclusion hsa_circ_0044569 is highly expressed in lung cancer and related to the pathological type, suggesting that it may become a potential biomarker in the early diagnosis of lung cancer.
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Objective:To investigate the expression of hsa_circ_0019413 in the peripheral blood of patients with primary Sj?gren's syndrome (pSS) and its role in the development of pSS disease.Methods:Microarray screening of circ ribonucleic acid (circRNA) changes was first performed in the peripheral blood of 4 pSS patients and 4 healthy controls. Real-time quantitative reverse transcription polymerase chain reaction (qPCR) was used to verify the difference in the expression of hsa_circ_0019413 in the peripheral blood of 30 pSS patients and 30 controls. By establishing the receiver operating characteristic (ROC) curve, the potential diagnostic value of hsa_circ_0019413 in peripheral blood was analyzed, and the expression level of hsa_circ_0019413 was correlated with the clinical presentations of patients with pSS.Results:① By microarray analysis, 437 circRNAs were differentially expressed between the two groups (FC≥2.0, P<0.05), of which 365 were up-regulated and 72 were down-regulated. ② The expression level of hsa_circ_0019413 in pSS patients was significantly higher than that in healthy controls by qPCR. The difference between the two groups was statistically significant ( P<0.05). It showed that hsa_circ_0019413 in peripheral blood of pSS patients had potential diagnostic value by ROC curve analysis [area under the curve (AUC)=0.883, 95% CI (0.782, 0.984), P<0.01]. ③ The expression level of hsa_circ_0019413 was positively correlated with the ESSDAI, ANA, titer of the pSS patients by correlation analysis ( r=0.721, P=0.012; r=0.625, P=0.040), but not with (immunoglobulin (Ig)G or erythrocyte sedimentation rate (ESR). Conclusion:Hsa_circ_0019413 in the peripheral blood may be involved in the development of pSS and may be a biomarker for the diagnosis of pSS.
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Objective To study the association of the CAG repeat length polymorphism in exon 1 of the androgen receptor (AR) gene with metabolic syndrome (MS) and its components in men.Methods We used cluster sampling method to select 910 middle-and old-aged male subjects from the communities in Yinchuan and Wuzhong cities,Ningxia.Their body height,weight and blood pressure were measured;their testosterone (TT),serum lipids,fasting glucose (FBG) and fasting insulin (FINS) were assayed;and their body mass index (BMI) and free testosterone (FT) were calculated.Length of CAG repeats in exon 1 of the AR gene was determined by polymerase chain reaction (PCR) and direct sequencing method.The subjects were divided into MS group (n =304) and normal control group (n=606) according to the diagnosis standards of MS.Results No obvious difference in the frequency distribution of CAG repeats in the AR gene was found between MS group and normal control group.The systolic blood pressure in the mcn with CAG repeat number of less than 22 was significantly higher than that in those with CAG repeat number of 22 or more,but HDL-C was significantly lower than men with CAG repeat number of 22 or more.However,the two groups did not significantly differ in diastolic blood pressure,other blood lipids level,FBG,FINS,BMI,TT or FT.The men with CAG repeat number of less than 22 had a higher prevalence of hypertension than those with CAG repeat number of 22 or more,but the prevalence of MS and other components of MS in the two groups had no significant difference,Conclusion The length of CAG repeats in exon 1 of the AR gene is related to the occurrence of hypertension and the dccreased level of HDL-C,but not to the incidence of MS.The number (less than 22) of CAG repeats of the AR gene may be a genetic factor of the occurrence of hypertension and reduced level of HDL-C.
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Objective To study the distribution of rs2234693 and rs9340799 of estrogen receptor (ER)gene and the relationship between them and lipid metabolism in the Ningxia Hui group,China.Methods We used cluster sampling method to select 5 3 3 cases of Ningxia Yinchuan communities of Hui. SNP genotyping was performed using the Sequenom MassARRAY platform.Results ① The frequencies of the distribution of each genotype of rs2234693 in Ningxia Hui population were the same as those in Chinese Han and Dai,Americans, Europeans,and Japanese (P>0.05).The distribution of each genotype of rs9340799,especially AA genotype,was different from that in Europeans (P0.05).Conclusion ER genes distribution in the Hui nationality of Ningxia is significantly different from that in other races;rs2234693 has no obvious relationship with the occurrence of dyslipidemia while GG genotype in SNP rs9340799 significantly increases the risk of lipid metabolism disorders.
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We developed a detailed electroporation method to deliver efficiently siRNA into mouse preimplantation embryos. By introducing Cy3 labeled negative control small interfering RNA (siRNA) into mouse preimplantation embryos, we optimized conditions for the electroporation, including the voltage, pulse duration, pulse number, electroporation buffer and an important step to weaken the zona pellucida. Embryonic survival rate, transfection rate and blastocyst development rate were evaluated under the converted fluorescence microscope, by embryos counting and statistical analysis. The best transfection was achieved in opti-MEM under the conditions of 30 V, 1 ms, 3 pulses, and the duration of digestion in tyrode's solution was 10 s. In conclusion, the proposed electroporation approach here is a simple and efficient tool to deliver siRNA for RNA interference (RNAi) into mouse preimplantation embryos.