Depiction of Vaginal Microbiota in Women With High-Risk Human Papillomavirus Infection.

Persistent infection with the carcinogenic human papillomavirus (HPV) is a prerequisite for the progression of cervical lesions and cancer. A growing body of research has focused on the functional role of the vaginal microbiota in the persistence of HPV infection. Understanding the microbial composition and structure in women with high-risk (hr)-HPV infection may help reveal associations between the vaginal microbiota and HPV infection, and identify potential biomarkers. Our study investigated the vaginal microbial community in women with and without hr-HPV infection, by using 16s rRNA gene sequencing. We found that microbial perturbations occurred in the early phase of hr-HPV infection. Lactobacillus and Sporolactobacillus were decreased, while bacteria related to bacterial vaginosis (BV), such as Gardnerella, Prevotella, Dialister, Slackia, Actinomyces, Porphyromonas, Peptoniphilus, Anaerococcus, Peptostreptococcus, Streptococcus, Ureaplasma, Megasphaera, and Mycoplasma were increased. Our results could offer insights into the correlations between hr-HPV and the vaginal microbiota in the early infection period, and provide indications that the predominance of some BV-associated bacteria during hr-HPV infection may increase the risk for cervical neoplasia.

Characteristics of primary side population cervical cancer cells.

The aim of the present study was to identify and characterize side population (SP) cells in primary cervical cancer. A primary culture was successfully established, and the SP cells were isolated via fluorescence-activated cell sorting. Subsequently, in vitro analysis of clonogenic capacity by soft agar assay and in vivo analysis of tumorigenicity were performed. The isolated SP cells accounted for ~4.73% of the total primary culture cells. The SP cells had a decreased proliferation rate and an increased distribution in G0/G1 compared with non-SP (NSP) cells. Following isolation, SP cells exhibited increased proliferative and self-renewal potency compared with NSP cells. Furthermore, significant ATP binding cassette subfamily G member 2 (ABCG2) expression was detected in SP cells but not in NSP cells. The tumor formation rate of SP cells was longer, and the tumor size and tumor formation rate of SP cells were increased in non-obese diabetic/severe combined immunodeficiency mice. In conclusion, the present study demonstrated that SP cells can be isolated from primary cervical cancer cell culture, and SP cells are enriched with stem cell-like cells that have a high capacity for colony formation and tumorigenesis.

Y chromosome azoospermia factor region microdeletions and transmission characteristics in azoospermic and severe oligozoospermic patients.

Spermatogenesis is an essential reproductive process that is regulated by many Y chromosome specific genes. Most of these genes are located in a specific region known as the azoospermia factor region (AZF) in the long arm of the human Y chromosome. AZF microdeletions are recognized as the most frequent structural chromosomal abnormalities and are the major cause of male infertility. Assisted reproductive techniques (ART) such as intra-cytoplasmic sperm injection (ICSI) and testicular sperm extraction (TESE) can overcome natural fertilization barriers and help a proportion of infertile couples produce children; however, these techniques increase the transmission risk of genetic defects. AZF microdeletions and their associated phenotypes in infertile males have been extensively studied, and different AZF microdeletion types have been identified by sequence-tagged site polymerase chain reaction (STS-PCR), suspension array technology (SAT) and array-comparative genomic hybridization (aCGH); however, each of these approaches has limitations that need to be overcome. Even though the transmission of AZF microdeletions has been reported worldwide, arguments correlating ART and the incidence of AZF microdeletions and explaining the occurrence of de novo deletions and expansion have not been resolved. Using the newest findings in the field, this review presents a systematic update concerning progress in understanding the functions of AZF regions and their associated genes, AZF microdeletions and their phenotypes and novel approaches for screening AZF microdeletions. Moreover, the transmission characteristics of AZF microdeletions and the future direction of research in the field will be specifically discussed.

Isolation and characterization of cancer stem cells from cervical cancer HeLa cells.

Cervical cancer is one of the most common gynecologic malignancies and poses a serious health problem worldwide. Identification and characterization of cervical cancer stem cells may facilitate the development of novel strategies for the treatment of advanced and metastatic cervical cancer. Breast cancer-resistance protein (Bcrp1)-positive cells were selected from a population of parent HeLa cells using flow cytometry. The invasion capacity of Bcrp1-positive and -negative cells was analyzed with a Boyden chamber invasion test. The tumorigenicity of these cells was determined by in vivo transplantation in non-obesity diabetes/severe combined immunodeficiency (NOD/SCID) mice. The Bcrp1-positive subpopulation accounted for about 7% of the parent HeLa cell population. The proliferative capacity of the Bcrp1-positive cells was greater than that of the Bcrp1-negative cells (P < 0.05). In the invasion assay, the Bcrp1-positive cells demonstrated a greater invasive capacity through the artificial basement membrane than their Bcrp1-negative counterparts. Following transplantation of 10(4) cells, only the Bcrp1-positive cells formed tumors in NOD/SCID mice. When 10(5) or 10(6) cells were transplanted, the tumor incidence and the tumor mass were greater in the Bcrp1-positive groups than those in the Bcrp1-negative groups (P < 0.05). The Bcrp1-positive subpopulation cervical cancer stem cells.

[Research of the characterization of Bcrp1(+) HeLa cells].

OBJECTIVE: To make sure whether Bcrp1 is the marker of cervical cancer stem-like cells or not by studying the characterization of Bcrp1(+) HeLa cells. METHODS: Immunofluorescence stained flow cytometry and electron microscope were used to sort and observe ultrastructures of Bcrp1(+) and Bcrp1(-) HeLa cells. Flow cytometry was used to identify the cycle and the rate of apoptosis with annexin V in two group cells. The expression of proliferating cell nuclear antigen (PCNA) and caspase-3 were tested using western blot method. RESULTS: (1) There were 7.1% Bcrp1(+) cells and 92.9% Bcrp1(-)cells in HeLa cells. Bcrp1(+) HeLa cells were large in size of nuclear and nucleoli are clear, and there were rich of cytomicrosome and rough endoplasmic reticulum. After sorted and cultured for 24, 48, 72 hours, the adhesion in Bcrp1(+) cells were 72.8%, 81.1%, 80.4%, respectively. While, they were 3.3%, 18.7%, 12.6% at each time for Bcrp1(-) cells (all P < 0.05). (2) There are more S phase cells in Bcrp1(+) cells than that in Bcrp1(-) cells (54.1% vs 21.1%, P < 0.05), while the percentage of G(0)/G(1) and G(2)/M in Bcrp1(-) cells were higher than those in Bcrp1(+) cells (53.0% vs 44.4%, 25.9% vs 1.5%; all P < 0.05). The rate of apoptosis in Bcrp1(+) cells was lower than that in Bcrp1(-) cells (0.2% vs 5.3%, P < 0.05). (3) The expression of PCNA in Bcrp1(+) cells was higher than that in Bcrp1(-) cells (3140 vs 2255, P < 0.05), while the expression of caspase-3 of Bcrp1(+) cells was lower than that in Bcrp1(-) cells (1970 vs 3551, P < 0.05). CONCLUSION: There are more vigor and ability of proliferation and lower rate of apoptosis in Bcrp1(+) HeLa cells than those in Bcrp1(-) cells, which may be some characters of cervical cancer stem cells.