RESUMO
Aberrant DNA replication is one of the driving forces behind oncogenesis. Furthermore, minichromosome maintenance complex component 3 (MCM3) serves an essential role in DNA replication. Therefore, in the present study, the diagnostic and prognostic value of MCM3 and its interacting proteins in hepatocellular carcinoma (HCC) were investigated. By utilizing The Cancer Genome Atlas (TCGA) database, global MCM3 mRNA levels were assessed in HCC and normal liver tissues. Its effects were further analyzed by reverse transcription-quantitative PCR (RT-qPCR), western blotting and immunohistochemistry in 78 paired HCC and adjacent tissues. Functional and pathway enrichment analyses were performed using the Search Tool for the Retrieval of Interacting Genes database. The expression levels of proteins that interact with MCM3 were also analyzed using the TCGA database and RT-qPCR. Finally, algorithms combining receiver operating characteristic (ROC) curves were constructed using binary logistic regression using the TCGA results. Increased MCM3 mRNA expression with high α-fetoprotein levels and advanced Edmondson-Steiner grade were found to be characteristic of HCC. Survival analysis revealed that high MCM3 expression was associated with poor outcomes in patients with HCC. In addition, MCM3 protein expression was associated with increased tumor invasion in HCC tissues. MCM3 and its interacting proteins were found to be primarily involved in DNA replication, cell cycle and a number of binding processes. Algorithms combining ROCs of MCM3 and its interacting proteins were found to have improved HCC diagnosis ability compared with MCM3 and other individual diagnostic markers. In conclusion, MCM3 appears to be a promising diagnostic biomarker for HCC. Additionally, the present study provides a basis for the multi-gene diagnosis of HCC using MCM3.
RESUMO
OBJECTIVE: To evaluate the plasma membrane integrity and morphology of fresh and frozen goat spermatozoa. METHODS: The ejaculates of three male goats were obtained by the artificial vagina method of collection and the rates of sperm abnormality and acrosome integrity were detected after freezing-thawing processing. The plasma membrane integrity of the fresh and frozen-thawed goat spermatozoa was evaluated with a combination of fluorescent probes, carboxyfluorescein diacetate and propidium iodide. RESULTS: The freezing-thawing process significantly influenced the viability and integrity of the spermatozoa ([74.43 +/- 13.78]% vs. [46.25 +/- 2.69]%; [64.26 +/- 7.03]% vs. [6.27 +/- 2.90]%, P < 0.01). The results showed differences in acrosome integrity rate between the fresh and frozen samples ([80.77 +/- 10.70]% vs. [58.42 +/- 18.05]% , P < 0.05). CONCLUSION: The freezing-thawing process significantly reduces sperm viability and acrosome integrity and seriously damages the plasma membrane integrity.