Study On The Anti-tumor Mechanism Of IHZ-1 And Cycloastragenol In Preventing Tumors And Aging

Hepatocellular carcinoma(HCC)is the third highest mortality rate cancer in the world.The morbidity and morbidity gradually rise year after year.Nowadays,chemotherapy is still the main therapy of HCC.However,many anti-cancer compounds are not tissue specific and have toxic side effects.It is urgently need to develop high-efficiency and low-toxicity drugs.Our previous study found that IHZ-1 could inhibit the PI3K/Akt/m TOR signaling pathway and activate the ROS/JNK pathway to induce cell apoptosis in HCC cells.It well known that ROS plays an important role in apoptosis and autophagy.Autophagy is identified as a “self-eating” process which has a dual role including pro-death or pro-survival.The biological effect induced by autophagy is depending on the cell type and the level of stress.Therefore,we further investigated weather IHZ-1 can activate autophagy.We first used acridine orange staining to detect the number of acidic organelles.The results showed that the number of acidic organelles increased in HCC cells.Then,we further tested the m RNA expression of autophagy-related genes.The results showed that the transcription level of Beclin1,Atg5 and ULK1 are significantly increased after IHZ-1 treatment.In addition,the immunofluorescence and western blotting experiment both showed that autophagy marker LC3-II was increased after IHZ-1 treatment.However,the expression level of P62 which was a main autophagy substrate had no significantly change,suggesting IHZ-1 might be a autophagy inhibitor.To further confirm this result,we detected the expression of P62 using chloroquine(CQ)and IHZ-1 to co-treatment on HCC cells.The results showed that IHZ-1 significantly decreased the P62 expression in the presence of CQ.The above results indicate that IHZ-1 induced autophagy in HCC cells.Further study found that inhibition of autophagy increased the apoptosis ratio by IHZ-1.It suggested that the autophagy activated by IHZ-1 was a protective effect on HCC cells.In summary,IHZ-1 could accumulate ROS and induce autophagy in HCC cells,while inhibiting autophagy could enhance the apoptosis induced by IHZ-1.China has entered into the aging society with numerous age-related diseases,which seriously threatens people’s healthy life.More and more studies shows that the accumulation of senescent cells is an important mechanism to drive the occurrences and results in aging and aging-related diseases.The eliminating senescent cells or blocking the secretion of SASP are effective strategies for age-related diseases treatment.In 2011,it was the first time to reported that elimination of senescent cells from the body could prolong lifespan and delay age-related diseases.At the same time,further studies found that injecting senescent cells into young mice could decrease the lifespan up to 35%.Therefore,targeting senescent cells not only can prolong the lifespan,but also delay tumor formation and improve the organ function.However,the genetically modified models and gene therapy limite its practicality.Therefore,"senolytics" which is a class of compounds having selectivity pro-apoptotic effect on senescent cells becomes a new direction of anti-aging.Our previous study found that the cycloastragenol(CAG)could selectively eliminate senescent cells and inhibit SASP.The mechanism might be related to the down-regulation of Bcl-2 family,P38MAPK/NF-κB and JAK/STAT3.To further valua the anti-aging effects of the CAG in vivo,we established aging mice mode by D-gal treatment and intragastric administration of CAG.The result of body weight and organ weight ratio had no significantly change after CAG treatment.It suggested that CAG is not toxic.Then,we detected the bone density using Micro-CT and found that CAG rescued cortical bones decreasing induced by D-gal treatment.We also found that CAG slightly decreased the level of p16INK4 A in adipose tissue.Next,we used ELISA and Real-Time PCR to detect the expression level of SASP in serum and tissues.The results showed that CAG decrease the IL-6 and CXCL-10 expression.In addition,the expression of p-STAT3 also decreased in CAG treated mice.These results suggested that CAG could inhibit expression of SASP in vivo.In summary,CAG rescue partial aging phenotype of aging mice mode.The mechanism may be related to down-regulation of p16INK4 A and inhibit on SASP.