| Spectroscopy is an effective method in analyzing structure, component and content of substances due to its high sensitivity. It is an effective tool for studing the stages of the biomolecules and cells. X-rays is a kind of short wave electromagnetic waves, when the carcinomatous cells were irradiated by the X-rays, the protein and the nucleic acid will be damaged or repaired, at the same time, the X-rays irradiation will induce some mechanism of apoptosis. So X-rays is used to cure the malignant tumors in clinic. But to the clinic, there is no very scientific dose reference in the X-rays therapy, the design of the X-rays dose is always accoding to the experience of the doctors, it is very subjective.In this thesis, we collected the absorption spectra of the Hep-2 cells which irradiated by the different dose of X-rays and the culture medium (10%Foetal Calf Serum) cultivated the cells for 24h,48h and 72h respectively by spectrophotometer. On the other hand, the apoptosis of the Hep-2 cells which irradiated by the different dose of X-rays and cultivated for 24h was studied.Form thses outcomes, we tried to find out the rules of the interaction between the X-rays and the Hep-2 cells with the spectroscopy technoloGy, and tried to find out the best X-rays dose for the theropy of larynx carcinima to the clinic.This thesis includes four parts, and some important conclusions are drawn: in the 1st part, the influence to Hep-2 cells' UV-absorption spectra after the Hep-2 cells irradiated by X-rays and cultivated for 24h,48h and 72h was reported. It showed that,(1) there were visible differences among these absorption spectra of the Hep-2 cells irradiated by different dose of X-rays and cultivated for 24h,48h and 72h respectively, which related to not only the cell cycle but the irradiation of the X-rays,(2) the spctra at the time of 48h after the cells irradiated and cultivated is more characteristic, (3) the protein and the nucleic acid will be damaged and repaired induced by X-rays, which can be reflected from the spectra.In the 2nd part, Fourier transform infrared spectroscopy (FTIR) was used to study the Hep-2 cells irradiated by X-rays with different dose. Compared with the control group, the contents of protein, nucleic acid and fat in the higher dose groups were decreased obviously. And we found that the contents and frameworks of protein, nucleic acid and fat were changed most intensely in the 8Gy dose group. All of the results may offer some experimental evidences in search of the optimum irradiational dose of X-ray.In the 3rd part, the UV-absorption spectra of the Hep-2 cell's culture medium(10% Foetal Calf Serum ) after the Hep-2 cell irradiated by X-rays and cultivated for 24h,48h and 72h respectively were collected. From the results we found that there were visible differences among these absorption spectra. It showed that the content of each amino acid has already changed. That is, during the growing course of cultivating cancer cells, the tryptophan and casein were not depleted equivalently. And there were some important relations between the absorption spectra and the cells apoptosis and necrosis induced by X-rays. This will be a foundation for the best X-rays dose study for the larynx carcinoma.In the 4th part, the apopotosis was measured by the flow cytometry with Annexin V-FITC /PI; it showed that,the X-rays can induce the Hep-2 cells apoptosis obviously, the apoptosis relied on the X-rays dose but not linear.The apoptosis ratio of the 12Gy dose group was the highest, it was (20.793±0.133)%.But the highest of the apoptosis ratio of 1Gy dose (apoptosis ratio/dose) was the 2 Gy dose droup.The apoptosis of the dose group from 2Gy to 8Gy was chinged not markedly. |
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