Team:Shanghai-United/Description

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Inspiration:

Since HPV 9-valent vaccine officially entered the market in mainland China in April 2018, there has been immense difficulties in getting the vaccine. It was not only because of the high price, but also an apparent market shortage as so many people are waiting to get vaccinated that even making an appointment for the vaccine is like racing against a hundred other patients. Given the situation, some hospitals and institutions started to sell pseudovaccines. Several large cases have been filed, and the institutions were punished in the first half of 2019. This invoked our team's interest in the HPV vaccine. During our initial research phase, we found due to the highly limited number of HPV vaccines, most people who are most susceptible to HPV virus in developing countries did not get vaccinated. With the invention of HPV vaccine, the issue does not get assuaged. Our experiment was designed to find a new approach in diagnosing people with high risk of HPV in advance, so vaccines can be distributed in a most effective manner. Based on previous studies on the topic, we saw a consistent pattern of an unusual abundance of NF-X1 protein (a transcriptional factor) in HPV, cervical cancer patients. Thus, our experiment aimed to find the differential protein related to NF-X1, which could help with early diagnosis of HPV.

Description
All women are at risk of cervical cancer. It has become a leading cause of death of women. Although precancerous treatment can markedly lower mortality rates, they are still high worldwide due to limited access to cervical cancer screening. Here we demonstrate specific protein expression variations related to NFX1 over-expression, a gene found to be closely associated with cervical cancer.

During a transcription process, various types of transcription factors affect the ultimate gene expression. In general, they can be classified into two distinct groups: activators, which increment the transcription process, or repressors, which restrain transcription.

In MHC Class II gene transcription specifically, transcriptional factors binding to X and Y boxes in proximal promoter of the gene predominantly determine the gene's expression. The over-expression of NFX1-123 gene, which codes for a repressor protein binding to the X box, was found to be related with a higher risk of HPV infection.

However, particular pathways or mechanisms involved in this process remain unclear.

Here our team finds a list of proteins that are highly likely to be associated with an over-expression of NFX1-123.

Hela cells were either injected with empty vectors, or plasmids containing NFX1 and GFP genes. After they fully expressed inserted genes, we ran Western Blot, IP MS, and FASP to determine and confirm the differential proteins.

After careful analysis, the statistical outcomes of IP and FASP Mass Spectrum combined, indicated significant changes in the expression of 280 proteins caused by NFX1 overexpression. Our findings can serve as a convenient bio marker to identify people carrying over-expressed NFX1, signal high risk of cervical cancer, and prevent cervical cancer in early stages.

We found in controlled in vitro assays that an over expression of NFX1-123 results in different concentration of certain proteins compared to normal. Most importantly, our results are highly convincing because we utilized three distinct methods to confirm and filter the results, ensuring a strong relativity of NFX1-123 gene expression and the proteins found.

Our results indicate potential proteins engaged in HPV infection, providing a new method to diagnose people with high risk of HPV virus even before they are actually infected. Since cervical cancer can only be cured in early stages, this evaluation by protein concentration may serve as important signs that help us to prevent and combat cervical cancer well before it gets uncontrollable. We also anticipate our assay to be starting point for further research on the mechanisms of HPV virus. For example, a specific protein associated with NFX1 gene could be tested to examine the role of that protein in HPV infection.