The Synthesis Of E6/E7 Based CdTe Quantum Dots Nano Fluorescent Targeting Probe And Its Application In The Detection Of Cervical Carcinoma

Background and ObjectivesQuantum dots nano-fluorescent materials show unique chemical properties and photophysical properties, such as narrow and symmetric emission spectrum, wide and continuous excitation spectrum, strong light stability, long fluorescence lifetime, and are suitable in many aspects of biological detection, diagnosis and treatment, and tumor detection as fluorescence imaging materials. High risk-human papillomaviruses (HR-HPV) especially HPV16 are closely correlated with the development and evolution of cervical cancer. Sustained expression of HR-HPV16 E6/E7 is the key cause for immortalization of host cell, malignant transformation, malignant phenotype maintenance and invasion and metastasis, which means that E6/E7 targeted nano-fluorescent probe has the potential ability of recognition for HPVs related tumors. In this study, CdTe quantum dots were prepared in aqueous system and further coupled with E6/E7 antibody by EDC/NHS coupling reaction. Finally, HPV16 E6/E7 targeted nano-fluorescence imaging probe was obtained and used in fluorescence imaging for HPV16 E6/E7 positive cervical cancer CaSki cells.Methods1. Prepare CdTe quantum dots.2. Prepare HPV16 E6/E7 targeted nano-fluorescent probe.3. Culture HPV 16 E6/E7 positive cervical cancer CaSki cells as the experimental group, and HPV 16 E6/E7 negative C33A cells as the control group.4. In inverted fluorescence microscope, nano-fluorescent probe could recognize HPV 16 E6/E7 positive CaSki cells.Results1. Prepare CdTe quantum dots:in aqueous system and the final concentration of the prepared CdTe QDs was 10 mmol L-1. The average particle size of hydrated quantum dot particle size was 11.6nm. CdTe QDs showed strong fluorescence, which can be used for subsequent experiments.2. E6/E7 targeted nano-fluorescent probe was prepared through EDC/NHS coupling reaction with E6/E7 antibody3. The recognition ability of HPV16 E6/E7 targeted nano-fluorescent probe towards CaSki cells:incubated with nano-fluorescent probe for 12h, HPV16 E6/E7 positive CaSki cells (experimental group) showed fluorescence; incubated with nano-fluorescent probe for 12h, HPV16 E6/E7 negative C33A cells (control group) showed no fluorescence signal.4. The photostability of quantum dots:the cervical cancer CaSki cell incubated with CdTe quantum dots showed no photobleaching after 60 min of irradiation.5. Cell toxicity of quantum dots:it was found that the cultured cervical cancer CaSki cells showed no significant morphological changes after incubated with E6/E7 for 24 h.Conclusions1. HPV16 E6/E7 targeted CdTe nano-fluorescent probe was successfully prepared.2. HPV16 E6/E7 targeted CdTe nano-fluorescent probe could recognize specifically E6/E7 positive cervical cancer CaSki cell.