| Background:Oral squamous cell carcinoma (OSCC) is a kind of the most familiar malignant tumor in the mouth cavity tumor, have swollen80%of the mouth cavity’s cancer.The OSCC sufferer passed surgical operation and put to treat treatment effect in early days the contra was better, but for the sufferer in later period, though progress put to treat, chemotherapy and living creature treatment gave satisfaction for lord of after synthesis cured,5years, existing leading only for20%-40%, effect still incapability, How therefore to enhance the later period malignant tumor the treatment success ratio and patient’s survival quality is called the present stage domestic and foreign related research the hot spot.Nuclear factor-κB (NF-κB) is one kind has the multi-direction duplication control action nucleoalbumin factor, has the widespread biology activeness, after the activation, participates in many genes the duplication regulation, has the important meaning to cell’s survival.In ceases activity under the condition, NF-kappa B and NF-kappa the B inhibiting factor (inhibitor of NF-κB, IκB) unifies, exists by the non-active form in the cytoplasm. But gets what one deserves violently the system most important enzyme to be located at IκB the upstream IκB protein kinase compound IKK (IκB kinase, IKK). The cell after receiving the stimulation, IKK is activated the induction IκB degeneration, the activation NF-κB signal passage, the suppressor cell perishes weakly.IκBα-M is IκBα the gene mutant, inducts the cell it, the expression produces IκBα-M the protein to be possible with the p50-p65protein dimer union, but IκBα-M the variation protein cannot degrade by IκB the activating enzyme, therefore not IκB activating enzyme adjustment, thus blocks p65/NF-κB the signal conduction return route.Based on this, this experiment plans to Tca-8113the cell line cell Intermediate-range to enter IκBα-M the gene and to cause its expression, the attempt to block p65/NF-KB the signal conduction return route, induces the cell to perish weakly. And attempts the confirmation IκBα-M gene to have kills the cell function.Objectives:Inducts IκBα-M the gene to the tongue scale cancer cell, lights in the different time, NF-κB the signal transduction circuit Asia unit--p65protein expression degree as well as the cell perishes weakly through many kinds of method examination cell in the situation, and inquires into between both’s relevance. Thus obtains a series of images and the data information. Perishes weakly for the further gene induction tumour cell as well as the final clinical practice builds the good foundation.Methods:The experiment uses the person tongue squamose cell cancer Tca8113cell line, after the cell recovers, in37℃,5%CO2hatches in the box to raise. After treating the cell solstice logarithm vegetal period, through will be suitable the repeater with the lipin body2000IκBα-M the gene inducts in the cell. After the extension dyes, continues, in37℃,5%CO2hatches in the box to raise, and after the extension dyes different time (12h,24h,48h,72h) distinguish the application immunocytochemistry and Western Blotting detection signal, and simultaneously using the class type cell meter (Flow cytometry, after FCM) and the terminal nucleotide transferase lies between the biological element dUTP terminal mark law which leads (the TUNEL law) to examine the extension dyes, different monitoring team’s different time cell weak perishing rate.Results:1.Passing the immunity cell chemistry law can observe, the P65egg whiteses hardly appear in the cell nucleus, but is more been clampinged to the cell guilder in cycle of pit in the normal matched control cell in; After screwing and dying, the egg white begins mass wearing the super-afterbirth film and gradually enters region in the cell nucleus centre, this process at along with screw dye behind the increase in time but present increasing trend.2. draws support in western the blotting, P65protein in cell total protein content was discovered that may change gradually along with the time, it in the extension dyes in the group cell to express the quantity dyes after the extension the time to increase increases, in the extension dyes the latter48hours to achieve in the biggest other groups, the P65protein expression quantity changes not obviously.3. may discover after the analysis class type cell examination’s data:Lights in the different time, after the extension dyes, cell weak perishing rate changes unceasingly,48hours achieve again high. Other control group does not have the disparity.4. has the cell which perishes weakly in the TUNEL experiment to observe the more typical morphological change, like the chromosome enrichment, the nucleus disruption as well as perish weakly the minute and so on, perishes weakly the cell ratio change tendency and the class type cell examination result is the same.Conclusions:The extension dyes IκBα-M the gene, may induce the Tca-8113cell line the cell to perish weakly, after it weak perishing rate and the extension dye, the time is proportional, and weak perishing rate and the P65protein’s expression quantity become continuously the tendency. After the extension dyes, along with the time lapse, P65the branching location changes unceasingly in the space, at gathers finally with the cell nucleus around. The P65protein’s expression change and the spatial shift, perish weakly with the cell are being related. |
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