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Reprogramming of Human Cervical Cancer Cell (HeLa) in Mouse Preimplantation Embryo Microenvironment

Author: HaoZhenHua
Tutor: LiuHongLin
School: Nanjing Agricultural College
Course: Animal Genetic Breeding and Reproduction
Keywords: reprogramming cancer microenviroment aggregation blastocyst injection chimeras
CLC: R737.33
Type: Master's thesis
Year: 2007
Downloads: 70
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Abstract


It has been thought that malignant minor cells can invade various microenvironme-nt andwould cause teratocarcinoma after transplanting into adult mammals. But recent reportsshow that the metastatic phenotype of tumor cells can be reversed to normalcy after injectingto the blastocyst. There has been an increased focus on the role of tumor microenvironmentin cancer progression. In this study, three blastocyst microinjection, embryo aggregation andReal-time PCR were used for studying the effect of mouse preimplantation embryomicroenvironment on reprogramming of human cervical cancer cell.1 Production of aggregation chimerasCollect mouse eight-cell-stage embryos and morula and remove the zona pellucida. Placethe zona-free embryos individually into each of the depressions with 50-100 HeLa cells. AfterCulturinig for 30 hours, most of the aggregates were at the late morula or early blastocyst stag-e, the rate of blastocyst is 70.3%, 40 hours and 50 hours is 78.9% and 84.6%, respectively.2 Production of injection chimeras15-20 HeLa cells were introduced into one host embryo through blastocyst injection. Onlywell-expanded blastocysts should be used. Totally 97 chimeras were obtained and 73 embryosgot blastocyst cavity again after recovery culture of 24 hours. The hatching rate of embryos is75.3%.3 Expression of human cervical cancer cell susceptible genes in chimerasBased on the difference of theβ-actin cDNA sequence between human and mouse, thehuman special primers were designed by Oligo 6.0 for Realtime-PCR. The transcripts of humanβ-actin was detected in the aggregates by Real-time PCR, indicating that adult human cervical cancer cells can assemble with the mouse early embryos. After aggregation and injection,transcripts of human cervical cancer cell susceptible genes c-myc and c-erb B2 wassignificantly down-regulated (P<0.01).Our results demonstrate the ability adult humancervical cancer cells to respond to mouse embryonic environmental cues, a subset of which mayundergo a reprogramming of their malignant phenotype. This invitro model has the potential toprovide insights into the regulation of tumor cell by a preimplantation embryonic milieu, whichwill offer new idea for elucidating the intrinsic mechanisms and clinical therapy for cancer.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Uterine tumors
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