Proposed Implementation

Cervical cancer is the sixth most common female cancer in China, with both a high incidence and high mortality rate. In 2020, more than 600,000 new cervical cancer cases and 300,000 deaths were reported worldwide. According to statistics, it accounts for 80% of malignant tumors in the female reproductive system and shows a trend of increasing occurrence in younger patients. Despite technological advances in diagnostic cervical cancer testing techniques, the accurate diagnosis and stage determination of cervical cancer still relies heavily on the morphological assessment of cervical epithelial cells by experienced pathologists.nuclear-to-plasmicratio. However, this clinical case diagnostic assessment is usually subjective, which is an important reason behind the underestimation of the true incidence of cervical cancer[1]. In addition, it has been previously reported that some adjunctive imaging techniques for cervical cancer diagnosis are characterized by low sensitivity in cervical cancer, which creates the risk of potential missed diagnosis[2]. Therefore, the development of a new, highly accurate, and efficient screening technique is essential for the detection of cervical cancer in clinical practice. Our YiYe-China team aims to develop a “sensor” for cervical cancer screening, based upon the fluorescent variations of MALAT1-miRNA complexes in cervical cancer cells that can quantitatively reflect the disease’s progress in real-time, thus providing a powerful support for the early diagnosis of cervical cancer. This novel, efficient, and inexpensive detection method will pave the way for accurate diagnosis and clinical staging of cervical cancer, as well as revolutionizing a new intellectual approach to the subject.

Who are our proposed end users?

Since the phenotypes of cervical cancer are inconspicuous, and existed tools for its diagnosis are lack of accuracy and efficiency, so professional knowledge about cervical cancer is required. Misdiagnoses happen every year, causing missed the optimal treatment time. However, the sensor developed by our team uses HBC system, which can show changes of fluorescence clearly. The result allows ordinary doctors to diagnose correctly. In addition, the whole process is based on noncoding RNA and HBC system, and it doesn’t have any side-effect. Applying the tool, we only need to introduce recombinant plasmids that consist of LncRNA-MALAT1 and miR-145/ miR-22 into patients’ cells, and watch the expression of green fluorescent protein. Thus, this tool could be used to patients regardless of ages and health states.

At the same time, the cervical cancer sensor involves PCR, cell culturing, RNA recombination, and so on. Therefore, experimental training is needed and necessary. And when used in reality, doctors or nurses should ensure no mistakes because there is a cost for each time using the tool, and a single mistake may lead to severe dangers in patients’ health. The operators need to have appropriate certifications and must be able to use tools, such as pippeting, skillfully.

How do we envision others using our project?

Cervical cancer has a considerable curability at the early stage, while treatments become less effective along with the development of the cancer. Since testing tools nowadays are inaccurate, many females die from misdiagnosis. Fortunately, the situation will be changed by our team’s discovery, a sensor based on the abnormal expression of noncoding RNA in cells. Furthermore, equipment we used in the experiment is very basic, so that every formal laboratory could perform the procedure. This tool help diagnosing patients in early stage with much greater accuracy as well as a lower cost. Both factors contribute to the credibility of cervical cancer diagnosis, which encourages more people to check for cervical cancer in hospitals.

Moreover, on account of the comparatively simple procedure, most medical stuffs are able to practice it, and the diagnosis will be less time consuming. Patients no longer need to wait for a long time or make any preparation before the testing, so various inconveniences are avoided. In conclusion, we envision our tools for detecting cervical cancer being accurate, affordable, and convenient for all people. Thus, more patients could be discovered and take a therapy at early stages and be cured successfully.

How would we implement our project in the real world?

In order to apply our detection tool in the real world, we developed the skill in a cell-free system [3-4]. The system is able to mimic cellular activities in vivo, so we don’t need the step of transfection to practice our experiment. The process are divided into the following steps (figure 1):

Firstly, we extract cells directly from cervix and culture them into primary cell. Then, lysis of the cell pellet is extracted for fluorescence tests. Next, miR-22-sponge-pepper/miR-145-sponge-pepper plasmid are delivered into cervical cell extracts. After incubation for 2 hours, HBC530 dye are added into the mixture. Finally, operators could use fluorescence reader to observe the intensity of fluorescence expression.

According to our principles and results of experiments, a fluorescence change will be observed in cancerous cells, Hela cells. If a person is infected by HPV virus, there will be a low intensity of fluorescence in the person’s cells and a high intensity of fluorescence in his or her cells from control group. However, the variation degree is hard to define, so we need further clinical trials to find out that within what range can changes represent diagnosis of cervical cancer. In future, we plan to collect clinical data from healthy people and people with cervical cancer in different stages. Through quantitating fluorescent intensity, we can correspond brightness to numerical values, and create a standard based on data to determine the progress of the cancer by critical values.

Figure 1: the process of the experiment through cell-free system