Study On The Activity Of Aldosterone Reductase AKR1C Series Enzymes In Cancer Cells

The(AKR)superfamily of aldosterone reductase is a nicotinamide adenine dinucleotide phosphate dependent oxidoreductase.It has been found that four subtypes of AKR1 C are expressed in most human organs and are closely related to the occurrence,metastasis and drug resistance of cancer.At present,the relationship between the differential expression of AKR1 C in cancer cells in the same tissue and different tissues and tumorigenesis and drug resistance has been studied.In particular,whether it can reverse drug resistance by inhibiting the expression of AKR1 C has become a research hotspot in recent years.In this study,nine cell lines including hepatocyte HL-7702,hepatoma cell HepG2,cisplatin-resistant hepatoma cell HepG2/DDP,lung cell MRC-5,lung cancer cell A549,cisplatin-resistant lung cancer cell A549/DDP,breast cell MCF-10 A,breast cancer cell MCF-7 and cisplatin-resistant breast cancer cell MCF-7/DDP were selected.The expression of four AKR1 C enzymes in these three types and nine different cell lines was studied by real-time fluorescence quantitative PCR at mRNA level.Western Blot technique was used to study the expression of AKR1 C at protein level,transcriptome analysis was performed on liver,lung,breast cancer cells and corresponding drug-resistant cancer cells,and then the mechanism and pathway of drug resistance in drug-resistant cancer cells were studied.The four enzymes of AKR1 C in drug-resistant hepatoma cells,drug-resistant lung cancer cells and drug-resistant breast cancer cells were knocked down by liposome transfection,and the knock-down cells were treated in vitro to observe the reversal of drug resistance,and then to determine the action of AKR1 C in different drug-resistant cells.The results are as follows:1)At the transcriptional level,compared with normal cells,the expression of four AKR1 C enzymes was up-regulated in cancer cells and drug-resistant cancer cells.Hepatocellular carcinoma cells are compared with hepatocytes: AKR1C1 is 3.45 times,AKR1C2 is 2.97 times,AKR1C3 is 3.96 times,and AKR1C4 is 1.24 times;Comparison of drug-resistant liver cancer cells and liver cells: AKR1C1 is 1.473 times,AKR1C2 is 1.31 times,AKR1C3 is 5.358 times,and AKR1C4 is 0.306 times.Comparison of lung cancer cells and lung cells: AKR1C1 is 1.658 times,AKR1C2 is 1.37 times,AKR1C3 is 1.645 times,AKR1C4 is 1.25 times;drug resistant lung cancer cells and lung cells are compared: AKR1C1 is 5.69 times,AKR1C2 is 2.724 times,and AKR1C3 is 5.036 times,AKR1C4 is 5.905 times.Breast cancer cells compared to breast cells: AKR1C1 is 1.64 times,AKR1C2 is 1.221 times,AKR1C3 is 1.667 times,and AKR1C4 is 1.113 times;resistant breast cells are compared to breast cells: AKR1C1 is 4.336 times,AKR1C2 is 2.396 times,and AKR1C3 is 4.99 times,AKR1C4 is 1.28 times.2)At the protein level,compared with normal cells,the expression of four AKR1 C enzymes was up-regulated in cancer cells and drug-resistant cancer cells.The comparison between hepatoma cells and hepatocytes: AKR1C1 is 0.9122 times,AKR1C2 is 1.7914 times,AKR1C3 is 1.2943 times,AKR1C4 is 1.3511 times;drug resistant hepatoma cells and hepatocytes: AKR1C1 is 1.2854 times,AKR1C2 is 2.4836 times,AKR1C3 is 1.8652 times,AKR1C4 is 1.7301 times.The comparison between lung cancer cells and lung cells: AKR1C1 is 1.579 times,AKR1C2 is 1.1725 times,AKR1C3 is 0.9053 times,AKR1C4 is 0.8817 times;drug resistant lung cancer cells and lung cells: AKR1C1 is 2.0787 times,AKR1C2 is 0.914 times,AKR1C3 is 1.0276 times,AKR1C4 is 1.5042 times.Compared with breast cells,AKR1C1,AKR1C2,AKR1C3 and AKR1C4 were 1.2405,1.48,1.5194 and 1.5951 times respectively,while those of drug resistant breast cells were 1.6483,1.7969,2.1146 and 1.8184 times of AKR1C1,AKR1C2,AKR1C3 and AKR1C4 respectively.3)Transcriptome analysis showed that the AKR1 C family was slightly up-regulated in different cancer cells,which played a reducing role together,which was consistent with the trend of transcription and protein level.At the same time,it was found that the expression of cell membrane channel protein also increased significantly,which may be related to the drug resistance of cancer.4)Knocking down the four enzymes of the three kinds of drug-resistant cells respectively,and adding drugs to the knock-down cells in vitro,it was found that the reversal effect of inhibiting single protein on drug-resistant cells was not obvious,and the reversal effect of AKR1C3 in drug-resistant hepatoma cells was relatively obvious,the reversal effect of AKR1C4 in drug-resistant lung cancer cells was relatively obvious,and the reversal effect of AKR1C3 in drug-resistant breast cancer cells was relatively obvious,but the overall reversal multiple was not high,which was consistent with the results of the transcriptional group,indicating that AKR plays a role in the drug resistance of cancer cells by the whole family,not a protein,which is consistent with the results of the transcriptome.The results of this study provide experimental and theoretical basis for further study of the expression of four AKR1 C isozymes and AKR family in different cancer cells,as well as the occurrence,development and drug resistance of cancer cells,as well as the further development of cancer drugs and new cancer markers.