Effect Of IncRNA HOTTIP On Cervical Cancer Cell Proliferation, Migration And Invasion

Cervical cancer is a high incidence rate of cancer in young women. As the woman’s second popularlargest cancer, cervical cancer remains difficult for early diagnosis and poor of prognosis like other malignancies. Research has shown that HPV infection is presented in 99.8% of cervical cancer, however, HPV infection alone is not sufficient to induce cervical cancer. Gene mutation and abnormal transcription play important roles in this process, so it is particularly important to study the pathogenesis of cervical cancer.Long non-coding RNA （lncRNA）, which transcript is longer than 200nt, was originally considered as a genome transcriptional "noise". Recent studies have shown that, lncRNA is involved in silencing the X chromosome, genomic imprinting, chromatin modification, transcriptional activation, and transcriptional regulation. The expression levels of some lncRNAs are abnormal in colon cancer, breast cancer, liver cancer, etc. As described in the view of lncRNA related databases （lncRNA db, lncRNA disease database, etc.）.Some lncRNA such as BC200, MALAT1, MEG3, XLOC₀10588 and HOTAIR are associated with cervical cancer. It’s possible and necessary to explore more other lncRNAs destributing to the development of cervical cancer.Part I:HOTTIP expression is upregulated both in cervical cancer tissue and cancer cell lines.A sample panel comprised of 10 cases randomly selected from totally 50 cases is subjected to Human LncRNA Microarray V3.0 analysis, The results indicates that the expression levels of many lncRNAs are abnormal in cervical cancer tissues relative to adjacent normal tissues. Among which the expression level of lncRNA HOTTIP is significantly upregulated （6.3-fold） in cervical cancer as comparedpare to the corresponding adjacent tissues （P<0.01）, and in accordance with the tissue results, following q-PCR analysis reveals that the expression level of HOTTIP in cervical cancer cell lines Hela and C-33A is also significantly higher than that in normal epithelial cell line Hacat.Our results indicates that lncRNA HOTTIP is specifically upregulated in cervical cancer tissues and cancer cell lines.Part II:The functional effect of lncRNA HOTTIP on cervical cancer cell proliferation, migration and invasion.To investigate the potential biological functions of lncRNA HOTTIP on cervical cancer cell proliferation, migration and invasion. HOTTIP siRNA was transfected into Hela and C-33A cervical cancer cell lines, with negative siRNA as a control. Q-PCR assay was performed to confirm the knock-down of the level of HOTTIP. CCK8 assay and colony-forming unit assay were performed to evaluate the effect of HOTTIP knock-down on Hela and C-33A cell proliferation. Wound Healing assay was performed to valuate the effect of HOTTIP knock-down on Hela and C-33A cell proliferation and migration. Tumour invasion assay was used to evaluate the effect of HOTTIP knock-down on Hela and C-33 A cell invasion.Our study shows that the expression level of HOTTIP could be efficiently knocked down by siRNA strategy. And the HOTTIP knock-down could significantly suppress the tumor characteristics of cervical cancer cell, including the ability of proliferation, migration and invasion.In conclusion, the lncRNA HOTTIP is specifically upregulated both in cervical cancer tissue and cervical cancer cell lines. The HOTTIP knockdown could significantly suppress the tumor characteristics of cervical cancer cell, including the ability of proliferation, migration and invasion. Our results indicates that lncRNA HOTTIP may play important roles in the development of cervical cancer.