Over-expression Of Hsulf-1Increased The Cell Apoptosis In Lung Cancer Cell Lines A549and Analysis In Senile Patients With Lung Cancer

Objective: The aim of this study is to investigate the effect of the over-expressionof Hsulf-1on the proliferation and apoptosis of lung cancer A549cells by transfectiontechnology. This study provides experimental foundation for the possibility whetherHsulf-1could become a new therapy to the lung cancer. We analyze the cases to provewhether physical examination for the detection of lung cancer has clinical value.Methods: Synthetized Hsulf-1plasmid was transported into Lung cancer cellsA549by liposome（LipofectamineT M2000）.48hours after the transfection, we usedinverted phase contrast microscope to observe the cell morphological change. Thedifference of Hsulf-1mRNA expression was determined by reverse transcriptionpolymerase chain reaction（RT-PCR） and protein level of Hsulf-1was revealed bywestern blot analysis. Flow cytometry technology was used to detect the proliferationand apoptosis of cells. We compared the results of imaging report and pathologicalreport to discuss the veracity of radiographic examination in detecting lung cancer.Results: After transporting Hsulf-1plasmid into Lung cancer cells A549byliposome （LipofectamineT M2000） successfully, cells A549emerged inhibitedproliferation and increased apoptosis. Inverted phase contrast microscope was used toobserve the cell morphological change: Before transfection, cells grew well withadherence. They were fusiform or polygon and elongated the protuberances with roundcaryon and one or more nucleolus inside.48hours after the transfection, cells showedmorphological apoptotic change, poor to cling and increased number of floating cells.Cells became shrinkage, nuclear envelop breakdown and protuberances vanished. RT-PCR was used to detect the expression level of Hsulf-1mRNA and the resultshowed an increased level of Hsulf-1mRNA in the transfection group. Expressions ofblack control group and negative control group showed no statistically significantdifference compared to the pre-transfection. Plasmid with Hsulf-1gene could improvethe expression of Hsulf-1mRNA specifically. To detect the difference of Hsulf-1proteinlevel among three groups, we used Western Blot. The result as following: Hsulf-1protein level increased observably in the transfection group; expressions of Hsulf-1protein in black control group and negative control group showed no statisticallysignificant difference compared to the pre-transfection. From this result, we mightconclude that plasmid carrying Hsulf-1can induce the special upgrade of the protein ofHsulf-1. We used flow cytometry to detect the apoptosis rate and here were the results:the apoptosis rate of the transfection group was （19.53±1.32）%, while the number ofthe black control group was （0.58±0.21）%and the negative control group was （0.74±0.13）%. This data indicated that over-expressionof Hsulf-1can induce the apoptosisof cells A549.75%of the cases showed consistency of imaging report and pathologicalreport. Physical examination could be used as an important means of lung cancerdiscovered.Conclusion: Hsulf-1plasmid transported into human lung adenocarcinoma celllines A549to improve the expression of Hsulf-1can inhibit the growth and promote theapoptosis of the cell A549. Hsulf-1might become a new gene target to treat the lungcarcinoma. Patients with a nodule neuroimaging needed long term follow-up and mighthave a canceration possibility if the nodule progressed. Physical examination for earlydetection of lung cancer might have important significance.