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Diane Lefley

Research Technician, Faculty of Science

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Biography

I completed my undergratuate degree in Medical Materials Science and Enginnering at the University of Sheffield in 2003, from here I immediately started my first scientific position in the department of Oncology at the University of Sheffield as a junior technician.

I contributed to various projects, mainly bisphosphonate on breast cancer bone metastasis both in vitro and in vivo, within Oncology with Professor Ingunn Holen, and getting my MPhil with this group in 2009.

From here I started to work with different groups, first with Professor Gill Tozer in Oncology working on VEGF variants and then Dr. David Andrew in the Dental School working on Saracatinib as an analgesic.

I spent the next 5 years working with Dr. Penny Ottewell in Oncology, working on the effects og IL1B in breast cancer cells and metastasis both in vivo and in vitro, and developing in vivo models of bone metastasis.

I am currently working as a Research Technician in James Dixon's group in RMCT, BDI3.

Expertise Summary

In vivo techniques (murine):

Injection sc, iv, ip

Withdrawal/ injection ic

In vivo IVIS imaging

Minor surgery

In vitro techniques:

Cell Culture

IHC/ IF

ELISA

Western blots

rtPCR

CRISPR/ transfections

Simple flow cytometry

Past Research

2015-2021: The effect of IL1B on breast cancer cells.

Using CRISPR KO to generate IL1B or IL1R1 knock out cell lines, or lentivirus to generate IL1B or IL1R1 over expressing cell lines. Effect on proliferation, invasion and migration was assessed in vitro on MDA231 and MCF7 cells. Working alongside a post doc replicating in E0771 cells.

These cells were grown in vivo to assess their effect on metastasis using IVIS imaging, and in the case of E0771 cells, effect on the immune system ex vivo via IHC staining.

Assessing the synergistic effects of Anakinra (an IL1R anagonist) in combination with doxorubicin and zoledronaic acid (a treatment currently available for bone metastasis patients)

Develpment of a human tumour graft to human bone metastasis model. Using PDX (human xenografts) planted into the fat pad to metastasise to human bone chips from femoral head donations grafted sub cutaneously.

2013-2015: Saracatinib

Assessing the analgesic properties of saracatinib in combination with zoledronic acid on MRMT-1 cells grafted intra tibially into rodents. My role was to assess tumour size by IVIS imaging and by histology. Perform ELISAs on serum, maintain the cell line in tissue culture and perform western blots.

2012-2013: VEGF isoforms

Performing western blots and arrays to show the differences in proteins between the 3 isoforms of VEGF. Culturing cells with different VEGF isoforms in vitro, alongside fibroblasts and myocardiac cells. Immunofluorescent staining of subcutaneous tumours with different VEGF isoforms. IHC of subcutaneous tumours.

Performing intra venous injections of cells.

2003-2009: Bisphosphonates

Researching the effects of combining doxorubicin and zoledronic acid on cells in vitro and in vivo. Effects on proliferation, apoptosis, migration and accumulation of unprenylated Rap1a was assessed in vitro. Effects on tumours injected sc, intra tibially and by intra cardiac assessed in vivo. IHC on tumours to assess proliferation, apoptosis, macrophages, vessels and osteoclasts (in the bone).

MPhil on the effect of Risedronate analogues on MDA231 and MCF7 cells in vitro. Using proliferation, apoptosis and western blotting on 5 analogue compounds.

School of Pharmacy

University of Nottingham
University Park
Nottingham, NG7 2RD

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