The Mechanism Of Lactobacillus Through Notch-RBP Jk Pathway The Proliferation Of Hela Cells In Cervical Cancer

Objective:Cervical cancer is the most common gynecological malignant tumor,usually originates from the cervical intraepithelial neoplasia,99%is due to the type of high-risk human papilloma virus（HPV）HR-persistent infection caused by,however,is not a sufficient condition of cervical HPV infection,according to research shows that women at least once in a lifetime of HPV infection,but only a small percentage of the infection to continue to develop into cervical intraepithelial neoplasia,and cervical cancer.Lactic acid bacteria as the vagina,the advantage of the microbial flora can be simple sugars into lactic acid,maintain normal vaginal acid environment,produce the antimicrobial factor such as H₂O₂ and bacteriocin,thereby inhibit the growth of pathogenic microorganisms.At the same time,through the competitive exclusion mechanism,the pathogenic microorganisms are prevented from adhering to the vaginal epithelial cells,thus maintaining the microecological balance of the vagina.Studies have shown that lactobacillus is closely related to vaginitis,HPV infection,cervical intraepithelial neoplasia,the occurrence and development of cervical cancer.Notch pathway is a kind of signaling pathway with highly conserved coding genes,which is widely expressed in many species and play an important role in the differentiation of cells and the formation of tissue.It is found that the conduction abnormality of Notch signaling pathway is closely related to the occurrence of various human diseases,but the role of Notch in cervical cancer is still controversial.This experiment was to study whether lactobacilli by adjusting the expression of Notch-RBP Jk implementation to inhibit the action of the cervical cancer cell proliferation,for lactic acid bacteria in the prevention of cervical cancer and cervical cancer targeted therapy provides basis.Method:1Immunohistochemical experimentsSelect department received by the second affiliated hospital of Hebei Medical University between 2016.12 to 2017.9 total hysterectomy and cervical cancer radical tissue specimens,including of 20 patients with cervical squamous carcinoma specimens of normal cervical specimens in 5 cases,each specimen have complete clinical data.The expression of CSL and CyclinD1 in specimen was detected by immunohistochemistry.2The effect of lactobacillus on the Notch pathway in Hela cellsAt 5*10⁵/in every hole of 6 plates,culturing in 37℃,5%CO₂ incubator overnight.Lactobacillus acts on cells by MOI:1000.At the same time control group add 2 ml DMEM without antibiotics,cultivate for 12h,24h,36h,48h,60h,then collecting cells for testing the expression of CSL、CyclinD1 by Western blot.3After siRNA interferes with CSL expression in Hela cells,lactobacillus affects the Notch pathway in Hela cells3.1 Western blot detect siRNA interference effect3.2After siRNA interference on 8h,subsequently lactobacilli effect 48h,Western blot to detect the expression of CSL,CyclinD1 in protein level.4 Statistical methodsUsing SPSS statistical software（version 21.0）for data analysis.Result:1.Immunohistochemical HE staining showed cervical cancer tissue of squamous carcinoma,cervical squamous carcinoma tissues of CSL proportion is less than normal cervical tissue in the control group,the cervical squamous carcinoma tissues of CyclinD1 proportion is greater than the normal control group.2.Lactobacillus on Hela,Notch of transcription factor CSL protein expression level compared with blank control group increased significantly,and over time the higher expression,has the time dependence.3.The expression of CyclinD1 protein was significantly decreased after the effect of lactobacillus on Hela cells,and the decrease was obvious over time.4.After the expression of siRNA interference CSL,the expression of CyclinD1 in the Hela cells under the action of lactobacillus was significantly higher than that of the blank and negative group.It is suggested that CSL expression inhibits the expression of downstream target gene CyclinD1.Conclusion:1.The expression of CSL in cervical squamous cell carcinoma was less than that in normal cervical tissue,and the expression of CyclinD1 was more.2.The expression of CSL protein was significantly increased after the effect of lactobacillus on Hela cells3.The expression of CyclinD1 protein was significantly decreased after the effect of lactobacillus on Hela cells4.Lactobacillus inhibit the proliferation of Hela cells by up-regulating the expression of CSL in Notch pathway and inhibiting the expression of CyclinD1.