| Objective: Prostate cancer is one of the most common malignant tumors in men.Prostate cancer is more common in elderly patients.Due to the combined effect of various factors in the elderly,it is difficult to operate after prostate cancer,with more postoperative complications and poor prognosis.Therefore,it poses a certain threat to the recovery of elderly patients.Therefore,it is of great significance for prostate cancer to actively seek one or more effective drugs and methods to treat prostate cancer.At present,most of the treatments for cancer are to induce apoptosis of cancer cells by drugs to further anti-tumor.However,cell senescence has attracted much attention in anti-tumor research,as it can play an anti-tumor role by inhibiting cell proliferation.In China,natural compounds,as an important source of drugs,not only have small adverse reactions,low price,and not easy to develop drug resistance,but also have their unique advantages of anti-tumor,so they have a good development prospect,and become a hotspot in the field of medicine.Osthole is a natural coumarin compound,mainly extracted from the natural compound Osthole,which has anti-inflammatory,antioxidant,antipruritic,anti-asthma,antiosteoporosis,immune regulation,promoting fracture healing,anti-bacterial,anti-viral and anti-tumor pharmacological effects.It has been reported that cestin can induce apoptosis of a variety of cancer cells and achieve anti-tumor effect.About common cnidium fruit,however,induced the prostate cancer research is few,especially about the related research of aging rarely happened in induction of prostate cancer cells,this study by cells in vitro experimental study fructus cnidii element impact on prostate cancer LNCap cell senescence and its mechanism of action,in order to common cnidium fruit,new drug development and provide certain reference for clinical treatment.Methods: This experiment takes logarithmic phase of experiment,using the CCK8 experiment method,the concentration respectively(0,12.5,25,50,100,150 umol/L)of the common Osthole,role in prostate cancer LNCap cell 24,48,72 hours,to detect the influence of the cell vitality,and calculate the cells at different time points for(Half-inhibitory concentration50)IC50 alue concentration.The proliferation of LNCaP cells at different concentrations of Osthole was detected by EdU assay after 24 h of Osthole treatment,and the cycle of cell cycle arrest was further detected by flow cytometry after treatment with different concentrations of Osthole.The senescence of prostate cancer LNCap cells treated with different concentrations of Osthole for 24 hours was detected by the staining method of-galactosidase.After the staining,the cells were placed under an inverted light microscope for photos,and the morphological differences between senescent cells and normal cells were further observed and compared.Western Blot method was used to detect the expression of senescence related proteins Skp2 and P27 in LNCap cells of prostate cancer after 24 hours of continuous action with different concentrations of Osthole.Result: 1.Osthole inhibits the growth and proliferation of prostate cancer cells by blocking the cell growth cycle in G2/M phase: The results of CCK8 test showed that,after 24,48,and 72 hours of different concentrations of Osthole on LNCaP cells of prostate cancer,Osthole could inhibit the proliferation of LNCaP cells with a time-dependent and concentration-dependent effect(P<0.05).The results of EdU experiment showed that cniposin could significantly inhibit the proliferation of LNCaP cells(P<0.05),and the cell cycle detection results showed that LNCaP cells could arrest the cell cycle in G2/M phase 24 hours after the drug treatment(P < 0.01).2.Osthole can induce the aging of prostate cancer cells: Common cnidium fruit,LNCaP role in prostate cancer cells after 24 hours,beta galactose glucoside enzyme staining showed positive staining cells was increased,said its LNCaP cell aging samples change can induce prostate cancer,can be found with the increase of drug concentration at the same time,the aging of the cells was also increased,presents a certain concentration of correlation(P < 0.001).3.Osthole can cause changes in aging-related proteins in prostate cancer cells: The expression of protein Skp2 and P27 was detected by Western Blot: with the increase of drug concentration,the expression of protein P27 increased gradually,while the expression of Skp2 decreased gradually,with statistically significant difference(P<0.05).Conciusion:1.Osthole can not only reduce the activity of human prostate cancer LNCaP cells,but also block the cell cycle in G2/M phase,thus inhibiting the proliferation of the cells and showing a certain time dependence and concentration dependence;2.Osthole can induce senescence like changes in human prostate cancer LNCap cells with a certain concentration dependence;3.Osthole can induce senescence in prostate cancer LNCaP cells by regulating the expression of senescence related proteins,including down-regulating the expression of protein Skp2 and up-regulating the expression of protein P27. |
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