The Biological Behavior And Mechanism Of NDUFA4 In Pancreatic Cancer

OBJECTIVE:Pancreatic cancer will become a huge public health burden due to difficulties in early diagnosing and poor prognosis.New therapies such as immunotherapy are not effective enough in this area,and resistance to first-line chemotherapy is the core problem that needs to be solved urgently.Therefore,it is of great significance to find the mechanism of chemoresistance and get new targets.In the early stage of our research group,high-throughput transcriptome sequencing was used to screen out multiple genes with differential expression between the gemcitabine-sensitive group and the gemcitabine-resistant group,suggesting that it is related to chemoresistance,including NDUFA4,which may have an impact on the biological behavior of pancreatic cancer cells and the sensitivity of chemotherapy.NDUFA4 gene is reported to activate growth factors through Bcl-2 and cytochrome C-mediated signaling pathways to inhibit apoptosis and promote growth in neurons;it has also been shown to have regulatory effects in a variety of tumor cells,although the mechanism is still to be further explored.This research intends to explore whether inhibiting the expression of NDUFA4 under in vitro conditions will affect the proliferation,apoptosis,and chemotherapy sensitivity of pancreatic cancer cells.It will be the preliminary exploration of this potential regulator,further research of NDUFA4 is to be carry out to show the possible mechanism of chemoresistance and to evaluate its potential as a diagnostic marker or a therapeutic target.METHODS:Western blot was used to detect the expression level of NDUFA4 in different pancreatic cancer cell lines,and the tool cell lines with moderate expression were screened;Use siNDUFA4 to transfect tool cell lines,and the transfection efficiency was detected by real-time quantitative PCR;CCK-8 method and flow cytometry were used to detect the changes in cell proliferation and apoptosis after down-regulation of NDUFA4 expression;Different concentrations of gemcitabine treated stably transformed cell lines,CCK-8 method was used to detect the difference in chemotherapy sensitivity;Gemcitabine treated tool cell lines were extracted Total mRNA,real-time quantitative PCR to detect whether gemcitabine can induce differences in NDUFA4 expression.RESULTS:NDUFA4 is highly expressed in cells in the gemcitabine resistance group,and there are differences in expression in different types of pancreatic cancer cell lines;It is predicted that down-regulation of NDUFA4 expression will make the OD value of transfected cell line less than that of the control group,proving that down-regulation of NDUFA4 can inhibit proliferation of pancreatic cancer cells;Flow cytometry analysis predicts that down-regulation of NDUFA4 can inhibit the apoptosis of pancreatic cancer cells;After gemcitabine treatment,it is predicted that the inhibition rate of stable transfected cell lines is higher than that of the control group,proving that down-regulation of NDUFA4 can increase the sensitivity of chemotherapy;It is predicted that the level of NDUFA4-mRNA in the cell line after gemcitabine treatment is higher than that in the control group,proving that gemcitabine can induce upregulation of NDUFA4 expression.CONCLUSIONS:NDUFA4 gene is differentially expressed in different pancreatic cancer chemo-resistant cell lines;Down-regulation of NDUFA4 gene expression can inhibit the proliferation of pancreatic cancer cells and promote the process of apoptosis.Down-regulation of NDUFA4 increases the sensitivity of chemotherapy;Gemcitabine can induce up-regulation of NDUFA4 expression.This shows that NDUFA4 is one of the potential influencing factors of chemotherapy resistance in pancreatic cancer,and worth further research.