| High metastic tongue squamous cell carcinoma line(Tca-8113) are heated to 43°C in vitro. The cells are observed and analysis by invert microscope(LM), transmission electron microscope (TEM), cell adhesiveness ssay, immunohistochemistry, flow cytometry and zymography. To investi-gate the morphology alteration, ultra-structure, cell cycle, DNA content, cell growth activity , biology behavior, laminin receptor(LNR) and expres-sion of matrix metallo -proteinase(MMP-2, MMP-9, TIMP-2) of tongue squamous cell carcino -ma line heated. Try to find hyperthermia theory base and the mechanism of tumor cell inva -sion are suppressed, so that the molecul and pathology principle in which hyperthermia suppress tumor can be disclos -ed to provide theory base for clinical tumor therapy.Control group: tongue squamous carcinoma cells(Tca-8113), cultivated for 24 hours at 37centigrade. Experimental group: 1. After Tca-8113 cells are heated to 43 centigrade each for 10, 20,30,40 minutes, they are cultivated for 24 hours. The obtained specimens are dyed with Giemsa and observed by LM. Other specimens, heated for another 40 minutes, cultivated for another 0,3,6, 12,48,72 hours, are determined DNA,MMP-9,MMP-2,TIMP-2,LNR by flow cytometry. 2. Tca-8113 cells heated for 40 minutes, arecultivated for 24 hours. Then, the obtained specimens are analysed byTEM, cell adhesion, MMP-9^ MMP-2^ TIMP-2 expression qualities. 1. LM: The heated cells contracted, along with nuclear wrinkled. Cells alined sparsely. The cells slowed down their growth. 2. TEM: The control group cells are large, abnormal and large nuclear. In the cytosols, there are sparse organelles. few mitochondrion, endoplasmic reticulums and many ribosomes can be seen. The experimental group cells are large, abnormal nuclears, many mitochondrion, ribosomes, Golgi complexes, microfiloments can be seen in these cells. 3. Cell adhesion: The experimental group cells adhension to FN and LN is lower than the control group cells(P<0. 05). 4. Immunohistochemistry: MMP-2.MMP-9 and TIMP-2 are expressed in both groups,but MMP-2 and MMP-9 decreased expression after heated, however, TIMP-2 expression increased.5. Detection of LNR> MMP-2> MMP-9 and TIMP-2: In heated cells, the expression of LNR, MMP-2> MMP-9 decreased, TIMP-2 increased.6. DNA detection: Hyperthermia can' t effect Go-d. S phage cells, After heated 10 minutes, G2-M phage cells dramatically decreased, but this effect can' t change as the heated time prolonged. 7.Detection of vitalities of MMP-2. MMP-9 by zymography: In heated cells, the vitalities of MMP-2^ MMP-9 decreased.1. The cell membrane is wrinkly, present a great guantity vacuole and cellular organelle, and the cell growth slow down after heating tongue squamous carcinoma cell (Tca-8113). All indicate that the cell become more and more ripe, the different -tiation is better, the malignant phenotypes take a turn, the malignant degree isreduced, the invasiveness was suppressed. 2. The percentage of Go-Gi-. S period has not changed and the percentage of G2-M period descensus obviously after heating tongue squamous carcinoma cell. The findings indicate that hyperthermia is not influence normal growth, but can suppress the cell proliferation and invasive ness. 3. The cell adhersion to LN, FN and ECU are reduced after heating tongue squamous carcinoma cell (Tca-8113). It show that the cell invasiveness is reduced. 4. The expression of MMP-^ MMP-9 reduce and the expression of TIMP-2 increase after heating tongue squamous carcinoma cell (Tca-8113). It is suggested that hyperthermia can suppress tumor cell invasiveness through changing the expression and activity of MMPs, in order to achieve the goal of curing cancer.
|
PDF Full Text Request