MicroRNA Profiling in Cervical NeoplasmsmicroRNA Array Services
Bali Muralidhar is working in Dr Nick Coleman's group at the Medical Research Council Cancer Cell Unit in Cambridge, UK. The group is investigating novel approaches to cancer diagnosis.
This involves two main areas: 1) The development of novel markers for improved screening for cervical cancer and colorectal cancer, and 2) The mechanisms of cervical neoplastic progression.
1. What is the current research going on in your lab? Basically we are a Papilloma Virus research lab that is looking into cervical cancer. We’re looking for genes that could be responsible for progression of cervical carcinogenesis. We’re also looking for biomarkers of progressive disease that could be used in clinical practice.
2. What made you want to study microRNAs? We found that one of the major microRNA processing enzyme, Drosha, is a marker that we picked up in our W12 cervical carcinogenesis model as being important in the progression of cervical carcinogenesis through invasion and migration.
3. What is your previous experience in microRNA profiling? None! We have done a lot of array CGH in the lab, and a fair amount of gene expression arrays. But we had never done any microRNA profiling work at all, which is why we decided to use a service provider for our microRNA profiling studies.
4. Why did you choose to use the miRCURY LNA™ Array microRNA profiling services from Exiqon? The main reason we chose Exiqon was because of the LNA ™ technology. It seemed to be technically a much better system than was available on the market elsewhere, and the results that we could see on the Exiqon website seemed to be really good. We also spoke to people that had experience with other microRNA array platforms, and they suggested that we should use the Exiqon LNA ™ technology. We looked into the different options and decided to go with Exiqon in the end.
5. In your opinion what are the main advantages to using the miRCURY LNA™ Arrays compared to other types of microRNA array? Since submitting samples to Exiqon, we have also tried using a home-built DNA based array for microRNA profiling. We found the Exiqon LNA ™ array results to be much more reproducible, and we were able to validate the results by qRT-PCR1. When we ran some of the same samples using the home-build DNA based array, there was much higher background on the arrays, and we got different results! We were not able to validate the differentially expressed microRNAs by qRT-PCR, so the changes that we saw on the home-built DNA based array were probably not real.
6. What do you feel are the main advantages of using a service provider, instead of performing experiments in your own lab? Service providers have a lot more experience in running these types of experiments, all the experimental conditions are consistently maintained and the results are much more reproducible. After an initial few glitches, our experiments are working well.
7. How do you feel about the results you received from the microRNA profiling services? Really good. I like the layout of the final report of the results and I like the basic data analysis that is performed. I think the fact that Exiqon provides some bioinformatics is really valuable for researchers that may not have access to a bioinformatics facility themselves.
8. How did you go on to validate the results from the microRNA arrays? We used TaqMan microRNA assays and were able to validate the differentially expressed microRNAs identified using the Exiqon LNA ™ arrays 1 .
9. Currently, what is the biggest challenge in your microRNA research? Functional Analysis, definitely, and actually discovering potential targets of these microRNAs. It’s all well and good to find out that microRNA x or y changes with treatment z, but you really need to identify the real biologically relevant targets of those particular microRNAs.
1 Muralidhar B. et al. Global microRNA profiles in cervical squamous cell carcinoma depend on Drosha expression levels. J Pathol. 2007 Apr 30;212(4):368-377