| Background:Lung cancer is one of the most common malignant tumors over the world,which has become the number one killer of human being health.More than 1.6 million people die of lung cancer each year around the world.Owing to the high mortality rate,low survival rate and poor prognosis the lung cancer,not only affects people's quality of life,but also brings a heavy economic burden to families and society.In recent years,with the advancement of surgical treatment,radiotherapy,chemotherapy,immunotherapy,targeted therapy and other treatments,the prognosis of some patients has been improved.However,due to the difficulty in early diagnosis,a considerable number of patients have been found to be in the advanced or even end-stage,losing the opportunity of traditional treatment(such as surgery,radiotherapy and chemotherapy).Targeted therapy,as a hotspot in the field of cancer research in recent years,has shown great potential in the treatment of lung cancer,but due to various reasons such as drug resistance and lack of sensitive targeted genes,owing to these reasons,looking for new targets for patients become necessary.Experimental Objective:1.To study the biological function changes of non-small cell lung cancer cells expressing PAQR3 and knocking down PAQR3;2.To explore the molecular mechanism related to the function changes of cells.Experimental Methods:Observed the migration and invasion ability in cells that overexpression and knockdown of PAQR3 in non-small cell lung cancer cells by scratch and Transwell methods.Western blot detection of migration,invasion(EMT related E-cadherin and Vimentin)related indicators;2.Detection of cells Viability and cloning ability by colony formation and MTT assay;3.Flow cytometry was used to detect the death of A549 and H1299 cells,and the expression of apoptosis-related protein caspase3 was detected by western blot.4.Western blot was used to detect the expression of MEK-related proteins to explored the regulatory mechanism of PAQR3 on non-small cell lung cancer.Experimental Result:Overexpression of PAQR3 inhibited the migration and invasion of A549 and H1299.Knockdown of PAQR3 promoted the migration of non-small cell lung cancer cells,but did not promote its invasion,and EMT related protein have changed(with the PAQR3 overexpression the E-calcium increased,vimentin decreased;decreased expression of PAQR3,the E-cadherin decreased,vimentin expression increased);2.MTT assay showed that overexpression PAQR3 the cells viability was the lowest at the time of 48H;colony formation experiments showed that high expression of PAQR3 could inhibit the clonal ability of non-small cell lung cancer cells,but knocking down PAQR3 had no significant effect on the cloning ability of NSCLC cells;3.Flow cytometry experiment results showed that high expression of PAQR3 promoted apoptosis of A549 and H1299 cells,while knockdown of PAQR3 had no significant effect on cell apoptosis,and western blot showed that caspase3 was highly expressed in PAQR3 cells(especially with a molecular weight of 17KD).However,there was no significant difference between the caspase3 in the knockdown group and the control group.Western blot analysis showed that the expression of MEK signaling pathwayrelated proteins(Ras,Raf,ERK,p-ERK)in NSCLC cells was significantly decreased after overexpressiono of PAQR3,while the expression of MEK signaling pathway related proteins was slightly increased after knocking down PAQR3.Conclusion:PAQR3 is lowly expressed in non-small cell lung cancer.High expression of PAQR3 can inhibit the metastasis,invasio n,cloning and proliferation of non-small cell lung cancer cells by regulating MEK pathway,and promote the apoptosis of non-small cell lung cancer cells by regulating MEK pathway. |
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