Effects And Mechanism Of The TGF-? Signaling Regulator PMEPA1 On The Process Of Lung Cancer Metastasis To Bone

Lung cancer?breast cancer and prostate cancer are easily to metastasis to bone.And patients will suffer from severe osteodynia?pathological fracture?paraplegia?hypercalcemia and serious nerve compression which are called skeletal related events.On the end,patients will die.Nevertheless one of the most important causes is that cancer microenvironment and lung cancers have a series of functional interactions.Therefore,artificially cut off this interaction is a very effective treatment strategy,which can be designed a specific treatment to lung cancer metastasis to bone.TGF-? can regulate the expression of related genes that can promote the occurrence of bone metastasis.It shows that the TGF-?signaling pathway negative regulatory factor PMEPA1 plays an important role in the process of tumor cell EMT,and could inhibite prostate cancer metastasis to bone by inhibiting he TGF-?signaling pathway.In order to probe into the role of PMEPA1 gene in bone metastais of lung cancer,in this study,we selected two lung adenocarcinoma cell lines which were A549 and NCI-H292.The effects of TGF-?on the invasion?migration ability and related gene and protein expression of the two lung adenocarcinoma cells were observed before and after interfering the expression of PMEPA1 gene.Explore the effect and mechanism of PMEPA1 gene in the process of lung cancer metastasis to bone.ObjectiveWe used A549 and NCI-H292 as the research object,and tested the efficacy on the invasion?migration ability and the expression of related gene and protein after treated with TGF-?,before and after interfering PMEPA1 gene.Explore the effect and mechanism of PMEPA1 gene in the process of bone metastasis of lung cancer.Method1.RT-q PCR and Western blot method to detect the gene and protein PTHRP?MMP2 ? VEGF? IL-11 and ADAM19 which are related to invasion and migration ability,after treated with TGF-??2.Scratch test and Transwell to detect the effect of TGF-?on the invasion and migration of lung cancer cells.3.Using TGF-?to treat cancer cells after interfering PMEPA1 gene.4.RT-q PCR and Western blot to test the expression of related gene and protein.5.Scratch test and Transwell to detect the ability of invasion and migration.Results1.Result of scratch test: compared with the control couple,for 12 h the TGF-?couple migration distance ration : A549(181.53±11.34)%;H292(187.14±17.45)%.For 24 h,A549(177.32±8.14)%,H292(193.79±10.66)%.After interfering si RNA-NC and si RNA-PMEAPA1,for 12 h,A549(190.50±10.76)%?(283.10±33.00)%,H292(177.21±28.71)%?(279.24±16.61)%,for 24 h,549(201.67±35.12)%?(275.46±22.15)%,H292(162.05±12.53)%?(266.44±18.34)%?2.Result of Transwell invasion test:TGF-?enhances invasiveness of lung adenocarcinoma cells:A549(34±6.5)?(81±9.5);H202(52±7.5)?(122±6.1);after interfering si RNA,si RNA-NC and si RNA-PMEPA1:A549(82±5.0)?(162±7.85);H292(115±8.83)?(207±10.56)?3.Result of RT-qPCR:Expression of related genes compared to control couple after treatment with TGF-?%Expression of related genes compared to control couple after treatment with TGF-?%4.Western blotExpression of related proteins after treatment of TGF-?Expression of related proteins after treatment of TGF-?Conclusion1.TGF-?could enhance PMEPA1 gene and other genes and proteins which are related to bone metastasis of lung cancer.2.TGF-?can stimulate the invasion and migration ability of lung adenocarcinoma.3.TGF-?signal pathway negative regulatory factor PMEPA1 inhibits expression of bone metastasis related genes and proteins by inhibiting TGF-?signal pathway.