| Alcohol is considered to be an inducement for the formation and development of various tumors.Studies have shown that moderate and high alcohol consumption can increase cancer risk and promote cancer metastasis.There are few literatures about the effect of low alcohol intake on tumor cell metastasis,and most of them are epidemiological studies.The data of alcohol intake are obtained by questionnaire survey,so it is difficult to reach the same conclusion.Research conclusions on the relationship between alcohol consumption and tumors using animal cancer models are also inconsistent,mainly due to the different types of tumors and alcohol intake doses and methods.For example,most animal model studies use aqueous alcohol as the only drinking source for alcohol exposure It is impossible to accurately measure and control the dose of alcohol consumption in the experiment,especially for low alcohol intake.In summary,the effect of low concentration alcohol on tumor cell metastasis is still uncertain.The current method for evaluating the effect of alcohol on tumor cell metastasis is mainly to count lung metastases.Circulating tumor cells（CTCs）are the prerequisite and necessary way of tumor cell metastasis.By calculating the number of CTCs,we can predict the risk of tumor cell metastasis and the total survival time of patients.At present,there is no research work to investigate the effect of alcohol on the number of CTCs in animals.Therefore,the intake dose of alcohol is not accurate enough in epidemiological studies and animal model experiments.In this paper,the method based on the in vivo capture of circulating tumor cells is applied to the tumor model of mice with breast cancer,and the influence of different concentrations of alcohol on the mice tumor model CTCs is used to evaluate whether alcohol has an effect on the metastasis of breast cancer cells.In chapter one,tumor cell metastasis,the effects of ethanol on tumor cell metastasis,and the detection methods of CTCs are summarized.The effects of different concentrations of alcohol on tumor cell metastasis were introduced from two aspects of epidemiology and animal model.In chapter two,the quantity trend of CTCs captured in vivo was analyzed to evaluate the effect of alcohol on the metastasis of breast cancer cells.The effects of different ethanol concentrations,ethanol intake at different ages,ethanol intake time,and type of alcohol on breast cancer cell metastasis were investigated.At the same time of subcutaneous axillary inoculation of breast cancer cells,mice tumor models were perfused with alcohol of the corresponding concentration.On 7,14,21 and 28 days after inoculating,CTCs were captured,observed and counted by fluorescence staining.On day 35,mouse tumor models were dissected and lung metastases were counted.The results showed that different concentrations of alcohol had different effects on the breast cancer cell metastasis.The number of CTCs is consistent with the number of lung metastases.Ingestion of 1.0%w/v(4.38×10-2 g·kg-1)low concentration alcohol can significantly inhibit the breast cancer cell metastasis,intake of 20.0%w/v(0.878 g·kg-1)high concentration alcohol can significantly promote breast cancer cell metastasis,and could provide reliable basis for human drinking from the age,time and type of drinking.In chapter three,the current technology used for CTCs detection is only for certain specific types of carcinoma in situ,but alcohol can cause many types of tumors and clinical studies are usually uncertain about carcinoma in situ.In this chapter,a new method based on cocktail antibody in vivo capture and aptamer polychromatic fluorescence imaging was designed to detect two types of tumors,identify the in situ cancer of CTCs and study the role of alcohol in the unknown types of in situ cancer and its metastasis,so as to meet the needs of tumor diagnosis and clinical application..The antibody mixture of epithelial cell adhesion molecules（EpCAM）and epidermal growth factor receptor（EGFR）was used to capture breast cancer CTCs and liver cancer CTCs in vivo.Compared with EpCAM-based CTCs capture,the capture efficiency of CTCs in liver cancer increased significantly from 3.17%to 26.67%,and the capture efficiency of breast cancer CTCs increased slightly from 27.00%to 29.84%.At the same time,breast cancer CTCs and liver cancer CTCs were identified simultaneously by aptamer fluorescent probe staining without any signal crosstalk.The results of in vivo experiments with a double tumor mouse model confirmed the feasibility of this method in capturing CTCs and identifying cancer in situ. |
